US20220106549A1 - Cell culture cassettes and incubator - Google Patents
Cell culture cassettes and incubator Download PDFInfo
- Publication number
- US20220106549A1 US20220106549A1 US17/505,209 US202117505209A US2022106549A1 US 20220106549 A1 US20220106549 A1 US 20220106549A1 US 202117505209 A US202117505209 A US 202117505209A US 2022106549 A1 US2022106549 A1 US 2022106549A1
- Authority
- US
- United States
- Prior art keywords
- cell
- cells
- incubator
- cassettes
- cassette
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 238000004113 cell culture Methods 0.000 title claims description 43
- 238000000034 method Methods 0.000 claims abstract description 76
- 238000012258 culturing Methods 0.000 claims abstract description 20
- 230000008569 process Effects 0.000 claims abstract description 19
- 230000007613 environmental effect Effects 0.000 claims description 13
- 238000012544 monitoring process Methods 0.000 claims description 8
- 238000004891 communication Methods 0.000 claims description 5
- 238000003384 imaging method Methods 0.000 claims description 4
- 230000004044 response Effects 0.000 claims description 3
- 210000004027 cell Anatomy 0.000 description 145
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 63
- 201000010099 disease Diseases 0.000 description 60
- 230000000694 effects Effects 0.000 description 37
- 150000001875 compounds Chemical class 0.000 description 32
- 239000003795 chemical substances by application Substances 0.000 description 13
- 239000003550 marker Substances 0.000 description 11
- 230000001464 adherent effect Effects 0.000 description 10
- 239000000463 material Substances 0.000 description 10
- 102000004169 proteins and genes Human genes 0.000 description 10
- 108090000623 proteins and genes Proteins 0.000 description 10
- 102000004127 Cytokines Human genes 0.000 description 8
- 108090000695 Cytokines Proteins 0.000 description 8
- 206010028980 Neoplasm Diseases 0.000 description 8
- 230000009471 action Effects 0.000 description 8
- 239000003153 chemical reaction reagent Substances 0.000 description 8
- 230000007423 decrease Effects 0.000 description 8
- 150000003384 small molecules Chemical class 0.000 description 8
- 208000024891 symptom Diseases 0.000 description 8
- -1 for example Polymers 0.000 description 7
- 239000002609 medium Substances 0.000 description 7
- 238000012360 testing method Methods 0.000 description 7
- 108091034117 Oligonucleotide Proteins 0.000 description 6
- 230000003247 decreasing effect Effects 0.000 description 6
- 238000001514 detection method Methods 0.000 description 6
- 239000000975 dye Substances 0.000 description 6
- 239000012530 fluid Substances 0.000 description 6
- 239000003446 ligand Substances 0.000 description 6
- 239000012528 membrane Substances 0.000 description 6
- 235000015097 nutrients Nutrition 0.000 description 6
- 210000001519 tissue Anatomy 0.000 description 6
- 241000282414 Homo sapiens Species 0.000 description 5
- 230000006870 function Effects 0.000 description 5
- 239000007789 gas Substances 0.000 description 5
- 108020004999 messenger RNA Proteins 0.000 description 5
- 108020004707 nucleic acids Proteins 0.000 description 5
- 102000039446 nucleic acids Human genes 0.000 description 5
- 150000007523 nucleic acids Chemical class 0.000 description 5
- 238000012545 processing Methods 0.000 description 5
- 102000004190 Enzymes Human genes 0.000 description 4
- 108090000790 Enzymes Proteins 0.000 description 4
- 241000124008 Mammalia Species 0.000 description 4
- 108020004711 Nucleic Acid Probes Proteins 0.000 description 4
- 229920003182 Surlyn® Polymers 0.000 description 4
- 239000003242 anti bacterial agent Substances 0.000 description 4
- 230000003115 biocidal effect Effects 0.000 description 4
- 238000011534 incubation Methods 0.000 description 4
- 229910052500 inorganic mineral Inorganic materials 0.000 description 4
- 239000011707 mineral Substances 0.000 description 4
- 239000002853 nucleic acid probe Substances 0.000 description 4
- 239000000126 substance Substances 0.000 description 4
- 229920000089 Cyclic olefin copolymer Polymers 0.000 description 3
- 201000004624 Dermatitis Diseases 0.000 description 3
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 3
- 239000004793 Polystyrene Substances 0.000 description 3
- 206010039491 Sarcoma Diseases 0.000 description 3
- 206010003246 arthritis Diseases 0.000 description 3
- 208000006673 asthma Diseases 0.000 description 3
- 201000011510 cancer Diseases 0.000 description 3
- 210000003850 cellular structure Anatomy 0.000 description 3
- 238000000576 coating method Methods 0.000 description 3
- 238000004590 computer program Methods 0.000 description 3
- 208000035475 disorder Diseases 0.000 description 3
- 239000003814 drug Substances 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- 239000000284 extract Substances 0.000 description 3
- 239000001963 growth medium Substances 0.000 description 3
- 238000002347 injection Methods 0.000 description 3
- 239000007924 injection Substances 0.000 description 3
- 239000007788 liquid Substances 0.000 description 3
- 239000000178 monomer Substances 0.000 description 3
- 230000003287 optical effect Effects 0.000 description 3
- 229920003229 poly(methyl methacrylate) Polymers 0.000 description 3
- 229920000642 polymer Polymers 0.000 description 3
- 239000004926 polymethyl methacrylate Substances 0.000 description 3
- 229920000098 polyolefin Polymers 0.000 description 3
- 229920002223 polystyrene Polymers 0.000 description 3
- 238000012216 screening Methods 0.000 description 3
- 230000001225 therapeutic effect Effects 0.000 description 3
- 208000036762 Acute promyelocytic leukaemia Diseases 0.000 description 2
- 208000032467 Aplastic anaemia Diseases 0.000 description 2
- 208000023275 Autoimmune disease Diseases 0.000 description 2
- 206010005949 Bone cancer Diseases 0.000 description 2
- 208000018084 Bone neoplasm Diseases 0.000 description 2
- 239000004713 Cyclic olefin copolymer Substances 0.000 description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- 102000018997 Growth Hormone Human genes 0.000 description 2
- 108010051696 Growth Hormone Proteins 0.000 description 2
- 102000004195 Isomerases Human genes 0.000 description 2
- 108090000769 Isomerases Proteins 0.000 description 2
- 206010024612 Lipoma Diseases 0.000 description 2
- 208000019693 Lung disease Diseases 0.000 description 2
- 206010025323 Lymphomas Diseases 0.000 description 2
- 206010029260 Neuroblastoma Diseases 0.000 description 2
- 239000002033 PVDF binder Substances 0.000 description 2
- 102000035195 Peptidases Human genes 0.000 description 2
- 108091005804 Peptidases Proteins 0.000 description 2
- 208000033826 Promyelocytic Acute Leukemia Diseases 0.000 description 2
- 239000004365 Protease Substances 0.000 description 2
- 201000004681 Psoriasis Diseases 0.000 description 2
- 208000021386 Sjogren Syndrome Diseases 0.000 description 2
- 206010046851 Uveitis Diseases 0.000 description 2
- 239000000074 antisense oligonucleotide Substances 0.000 description 2
- 238000012230 antisense oligonucleotides Methods 0.000 description 2
- 230000001640 apoptogenic effect Effects 0.000 description 2
- 208000010668 atopic eczema Diseases 0.000 description 2
- 210000004369 blood Anatomy 0.000 description 2
- 239000008280 blood Substances 0.000 description 2
- 210000004556 brain Anatomy 0.000 description 2
- 239000001913 cellulose Substances 0.000 description 2
- 229920002678 cellulose Polymers 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 230000001684 chronic effect Effects 0.000 description 2
- 239000011248 coating agent Substances 0.000 description 2
- 208000029742 colonic neoplasm Diseases 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 230000007515 enzymatic degradation Effects 0.000 description 2
- 239000003102 growth factor Substances 0.000 description 2
- 239000000122 growth hormone Substances 0.000 description 2
- 239000005556 hormone Substances 0.000 description 2
- 229940088597 hormone Drugs 0.000 description 2
- 230000002757 inflammatory effect Effects 0.000 description 2
- 238000012423 maintenance Methods 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 201000008968 osteosarcoma Diseases 0.000 description 2
- 229920000728 polyester Polymers 0.000 description 2
- 229920006254 polymer film Polymers 0.000 description 2
- 229920002981 polyvinylidene fluoride Polymers 0.000 description 2
- 230000000750 progressive effect Effects 0.000 description 2
- 208000005069 pulmonary fibrosis Diseases 0.000 description 2
- 238000005070 sampling Methods 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- 230000001954 sterilising effect Effects 0.000 description 2
- 229940124597 therapeutic agent Drugs 0.000 description 2
- 238000012546 transfer Methods 0.000 description 2
- 230000003612 virological effect Effects 0.000 description 2
- 208000031261 Acute myeloid leukaemia Diseases 0.000 description 1
- 206010001052 Acute respiratory distress syndrome Diseases 0.000 description 1
- 208000022309 Alcoholic Liver disease Diseases 0.000 description 1
- 208000011403 Alexander disease Diseases 0.000 description 1
- 208000024827 Alzheimer disease Diseases 0.000 description 1
- 201000003076 Angiosarcoma Diseases 0.000 description 1
- 206010002556 Ankylosing Spondylitis Diseases 0.000 description 1
- 206010003399 Arthropod bite Diseases 0.000 description 1
- 102000014461 Ataxins Human genes 0.000 description 1
- 108010078286 Ataxins Proteins 0.000 description 1
- 208000032116 Autoimmune Experimental Encephalomyelitis Diseases 0.000 description 1
- 208000032791 BCR-ABL1 positive chronic myelogenous leukemia Diseases 0.000 description 1
- 206010004146 Basal cell carcinoma Diseases 0.000 description 1
- 208000023328 Basedow disease Diseases 0.000 description 1
- 208000008439 Biliary Liver Cirrhosis Diseases 0.000 description 1
- 208000033222 Biliary cirrhosis primary Diseases 0.000 description 1
- 206010073106 Bone giant cell tumour malignant Diseases 0.000 description 1
- 241000283690 Bos taurus Species 0.000 description 1
- 208000003174 Brain Neoplasms Diseases 0.000 description 1
- 208000014644 Brain disease Diseases 0.000 description 1
- 208000026310 Breast neoplasm Diseases 0.000 description 1
- 208000003170 Bronchiolo-Alveolar Adenocarcinoma Diseases 0.000 description 1
- 241000283707 Capra Species 0.000 description 1
- 241000282693 Cercopithecidae Species 0.000 description 1
- 206010008025 Cerebellar ataxia Diseases 0.000 description 1
- 201000005262 Chondroma Diseases 0.000 description 1
- 208000005243 Chondrosarcoma Diseases 0.000 description 1
- 201000009047 Chordoma Diseases 0.000 description 1
- 206010008909 Chronic Hepatitis Diseases 0.000 description 1
- 208000015943 Coeliac disease Diseases 0.000 description 1
- 206010009900 Colitis ulcerative Diseases 0.000 description 1
- 206010010741 Conjunctivitis Diseases 0.000 description 1
- 241000699800 Cricetinae Species 0.000 description 1
- 208000011231 Crohn disease Diseases 0.000 description 1
- 201000003883 Cystic fibrosis Diseases 0.000 description 1
- 206010011891 Deafness neurosensory Diseases 0.000 description 1
- 206010012438 Dermatitis atopic Diseases 0.000 description 1
- 206010012442 Dermatitis contact Diseases 0.000 description 1
- 208000019872 Drug Eruptions Diseases 0.000 description 1
- 208000003556 Dry Eye Syndromes Diseases 0.000 description 1
- 206010013774 Dry eye Diseases 0.000 description 1
- 229930193152 Dynemicin Natural products 0.000 description 1
- 208000000471 Dysplastic Nevus Syndrome Diseases 0.000 description 1
- 206010014561 Emphysema Diseases 0.000 description 1
- 208000032274 Encephalopathy Diseases 0.000 description 1
- 241000283073 Equus caballus Species 0.000 description 1
- 208000006168 Ewing Sarcoma Diseases 0.000 description 1
- 201000008808 Fibrosarcoma Diseases 0.000 description 1
- 206010016654 Fibrosis Diseases 0.000 description 1
- 206010053717 Fibrous histiocytoma Diseases 0.000 description 1
- 201000011240 Frontotemporal dementia Diseases 0.000 description 1
- 206010017993 Gastrointestinal neoplasms Diseases 0.000 description 1
- 208000007569 Giant Cell Tumors Diseases 0.000 description 1
- 206010018364 Glomerulonephritis Diseases 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- 201000005569 Gout Diseases 0.000 description 1
- 206010018634 Gouty Arthritis Diseases 0.000 description 1
- 206010072579 Granulomatosis with polyangiitis Diseases 0.000 description 1
- 208000003084 Graves Ophthalmopathy Diseases 0.000 description 1
- 208000015023 Graves' disease Diseases 0.000 description 1
- 208000035895 Guillain-Barré syndrome Diseases 0.000 description 1
- 208000002927 Hamartoma Diseases 0.000 description 1
- 208000030836 Hashimoto thyroiditis Diseases 0.000 description 1
- 208000001258 Hemangiosarcoma Diseases 0.000 description 1
- 206010019629 Hepatic adenoma Diseases 0.000 description 1
- 206010019755 Hepatitis chronic active Diseases 0.000 description 1
- 208000002972 Hepatolenticular Degeneration Diseases 0.000 description 1
- 208000017604 Hodgkin disease Diseases 0.000 description 1
- 208000010747 Hodgkins lymphoma Diseases 0.000 description 1
- 208000023105 Huntington disease Diseases 0.000 description 1
- 206010021245 Idiopathic thrombocytopenic purpura Diseases 0.000 description 1
- 208000004575 Infectious Arthritis Diseases 0.000 description 1
- 208000022559 Inflammatory bowel disease Diseases 0.000 description 1
- 208000029523 Interstitial Lung disease Diseases 0.000 description 1
- 208000003456 Juvenile Arthritis Diseases 0.000 description 1
- 206010059176 Juvenile idiopathic arthritis Diseases 0.000 description 1
- 208000002260 Keloid Diseases 0.000 description 1
- 208000009319 Keratoconjunctivitis Sicca Diseases 0.000 description 1
- 206010024229 Leprosy Diseases 0.000 description 1
- 208000009829 Lewy Body Disease Diseases 0.000 description 1
- 201000002832 Lewy body dementia Diseases 0.000 description 1
- 208000002404 Liver Cell Adenoma Diseases 0.000 description 1
- 206010058467 Lung neoplasm malignant Diseases 0.000 description 1
- 208000006644 Malignant Fibrous Histiocytoma Diseases 0.000 description 1
- 206010027406 Mesothelioma Diseases 0.000 description 1
- 206010049567 Miller Fisher syndrome Diseases 0.000 description 1
- 208000034578 Multiple myelomas Diseases 0.000 description 1
- 241000699666 Mus <mouse, genus> Species 0.000 description 1
- 208000015914 Non-Hodgkin lymphomas Diseases 0.000 description 1
- 239000004677 Nylon Substances 0.000 description 1
- 241000283973 Oryctolagus cuniculus Species 0.000 description 1
- 206010033078 Otitis media Diseases 0.000 description 1
- 229930012538 Paclitaxel Natural products 0.000 description 1
- 208000018737 Parkinson disease Diseases 0.000 description 1
- 241001494479 Pecora Species 0.000 description 1
- 208000000609 Pick Disease of the Brain Diseases 0.000 description 1
- 206010035226 Plasma cell myeloma Diseases 0.000 description 1
- 206010065159 Polychondritis Diseases 0.000 description 1
- 239000004698 Polyethylene Substances 0.000 description 1
- 208000012654 Primary biliary cholangitis Diseases 0.000 description 1
- 208000024777 Prion disease Diseases 0.000 description 1
- 206010036774 Proctitis Diseases 0.000 description 1
- 201000001263 Psoriatic Arthritis Diseases 0.000 description 1
- 208000036824 Psoriatic arthropathy Diseases 0.000 description 1
- 241000700159 Rattus Species 0.000 description 1
- 208000033464 Reiter syndrome Diseases 0.000 description 1
- 206010063837 Reperfusion injury Diseases 0.000 description 1
- 208000013616 Respiratory Distress Syndrome Diseases 0.000 description 1
- 208000005678 Rhabdomyoma Diseases 0.000 description 1
- 206010039710 Scleroderma Diseases 0.000 description 1
- 208000009966 Sensorineural Hearing Loss Diseases 0.000 description 1
- 201000010001 Silicosis Diseases 0.000 description 1
- 208000000453 Skin Neoplasms Diseases 0.000 description 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 1
- 208000009415 Spinocerebellar Ataxias Diseases 0.000 description 1
- 201000002661 Spondylitis Diseases 0.000 description 1
- 206010042033 Stevens-Johnson syndrome Diseases 0.000 description 1
- 231100000168 Stevens-Johnson syndrome Toxicity 0.000 description 1
- 206010042496 Sunburn Diseases 0.000 description 1
- 241000282898 Sus scrofa Species 0.000 description 1
- 206010043276 Teratoma Diseases 0.000 description 1
- 206010043781 Thyroiditis chronic Diseases 0.000 description 1
- 206010048873 Traumatic arthritis Diseases 0.000 description 1
- 206010070517 Type 2 lepra reaction Diseases 0.000 description 1
- 201000006704 Ulcerative Colitis Diseases 0.000 description 1
- 208000015778 Undifferentiated pleomorphic sarcoma Diseases 0.000 description 1
- 206010046914 Vaginal infection Diseases 0.000 description 1
- 201000008100 Vaginitis Diseases 0.000 description 1
- 208000018839 Wilson disease Diseases 0.000 description 1
- 208000027418 Wounds and injury Diseases 0.000 description 1
- JLCPHMBAVCMARE-UHFFFAOYSA-N [3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-[[3-[[3-[[3-[[3-[[3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-hydroxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methyl [5-(6-aminopurin-9-yl)-2-(hydroxymethyl)oxolan-3-yl] hydrogen phosphate Polymers Cc1cn(C2CC(OP(O)(=O)OCC3OC(CC3OP(O)(=O)OCC3OC(CC3O)n3cnc4c3nc(N)[nH]c4=O)n3cnc4c3nc(N)[nH]c4=O)C(COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3CO)n3cnc4c(N)ncnc34)n3ccc(N)nc3=O)n3cnc4c(N)ncnc34)n3ccc(N)nc3=O)n3ccc(N)nc3=O)n3ccc(N)nc3=O)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cc(C)c(=O)[nH]c3=O)n3cc(C)c(=O)[nH]c3=O)n3ccc(N)nc3=O)n3cc(C)c(=O)[nH]c3=O)n3cnc4c3nc(N)[nH]c4=O)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)O2)c(=O)[nH]c1=O JLCPHMBAVCMARE-UHFFFAOYSA-N 0.000 description 1
- 230000002378 acidificating effect Effects 0.000 description 1
- 230000001154 acute effect Effects 0.000 description 1
- 239000000853 adhesive Substances 0.000 description 1
- 230000001070 adhesive effect Effects 0.000 description 1
- 210000004100 adrenal gland Anatomy 0.000 description 1
- 201000000028 adult respiratory distress syndrome Diseases 0.000 description 1
- 201000009961 allergic asthma Diseases 0.000 description 1
- 208000026935 allergic disease Diseases 0.000 description 1
- 208000004631 alopecia areata Diseases 0.000 description 1
- 230000001668 ameliorated effect Effects 0.000 description 1
- 206010002022 amyloidosis Diseases 0.000 description 1
- 206010002026 amyotrophic lateral sclerosis Diseases 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 208000007502 anemia Diseases 0.000 description 1
- 230000000692 anti-sense effect Effects 0.000 description 1
- 208000002399 aphthous stomatitis Diseases 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 201000008937 atopic dermatitis Diseases 0.000 description 1
- 201000003710 autoimmune thrombocytopenic purpura Diseases 0.000 description 1
- 201000004982 autoimmune uveitis Diseases 0.000 description 1
- 201000004562 autosomal dominant cerebellar ataxia Diseases 0.000 description 1
- 208000001119 benign fibrous histiocytoma Diseases 0.000 description 1
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 1
- 230000002146 bilateral effect Effects 0.000 description 1
- 239000012472 biological sample Substances 0.000 description 1
- 230000005540 biological transmission Effects 0.000 description 1
- 239000007844 bleaching agent Substances 0.000 description 1
- 210000000988 bone and bone Anatomy 0.000 description 1
- 210000000481 breast Anatomy 0.000 description 1
- 239000011449 brick Substances 0.000 description 1
- 206010006451 bronchitis Diseases 0.000 description 1
- 208000003362 bronchogenic carcinoma Diseases 0.000 description 1
- 201000002143 bronchus adenoma Diseases 0.000 description 1
- 239000007975 buffered saline Substances 0.000 description 1
- 208000020670 canker sore Diseases 0.000 description 1
- 239000003990 capacitor Substances 0.000 description 1
- 239000011203 carbon fibre reinforced carbon Substances 0.000 description 1
- 239000000969 carrier Substances 0.000 description 1
- 230000021164 cell adhesion Effects 0.000 description 1
- 230000010261 cell growth Effects 0.000 description 1
- 230000003833 cell viability Effects 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 210000003679 cervix uteri Anatomy 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 238000007385 chemical modification Methods 0.000 description 1
- 208000006990 cholangiocarcinoma Diseases 0.000 description 1
- 201000005217 chondroblastoma Diseases 0.000 description 1
- 208000019069 chronic childhood arthritis Diseases 0.000 description 1
- 208000020832 chronic kidney disease Diseases 0.000 description 1
- 230000007882 cirrhosis Effects 0.000 description 1
- 208000019425 cirrhosis of liver Diseases 0.000 description 1
- 201000010897 colon adenocarcinoma Diseases 0.000 description 1
- 239000002131 composite material Substances 0.000 description 1
- 239000000470 constituent Substances 0.000 description 1
- 208000010247 contact dermatitis Diseases 0.000 description 1
- 239000000356 contaminant Substances 0.000 description 1
- 201000010305 cutaneous fibrous histiocytoma Diseases 0.000 description 1
- 208000004921 cutaneous lupus erythematosus Diseases 0.000 description 1
- 208000035250 cutaneous malignant susceptibility to 1 melanoma Diseases 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 239000008121 dextrose Substances 0.000 description 1
- 206010012601 diabetes mellitus Diseases 0.000 description 1
- 238000009792 diffusion process Methods 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 239000003937 drug carrier Substances 0.000 description 1
- 229920003247 engineering thermoplastic Polymers 0.000 description 1
- 210000003238 esophagus Anatomy 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 206010016629 fibroma Diseases 0.000 description 1
- 239000007850 fluorescent dye Substances 0.000 description 1
- 238000007756 gravure coating Methods 0.000 description 1
- 230000012010 growth Effects 0.000 description 1
- 208000024348 heart neoplasm Diseases 0.000 description 1
- 201000011066 hemangioma Diseases 0.000 description 1
- 201000005787 hematologic cancer Diseases 0.000 description 1
- 208000007475 hemolytic anemia Diseases 0.000 description 1
- 230000002008 hemorrhagic effect Effects 0.000 description 1
- 208000006359 hepatoblastoma Diseases 0.000 description 1
- 201000002735 hepatocellular adenoma Diseases 0.000 description 1
- 206010073071 hepatocellular carcinoma Diseases 0.000 description 1
- 230000002209 hydrophobic effect Effects 0.000 description 1
- 208000000509 infertility Diseases 0.000 description 1
- 230000036512 infertility Effects 0.000 description 1
- 208000021267 infertility disease Diseases 0.000 description 1
- 208000014674 injury Diseases 0.000 description 1
- 239000000976 ink Substances 0.000 description 1
- 201000004614 iritis Diseases 0.000 description 1
- 208000028867 ischemia Diseases 0.000 description 1
- 230000000302 ischemic effect Effects 0.000 description 1
- 201000002215 juvenile rheumatoid arthritis Diseases 0.000 description 1
- 210000001117 keloid Anatomy 0.000 description 1
- 206010023332 keratitis Diseases 0.000 description 1
- 201000010666 keratoconjunctivitis Diseases 0.000 description 1
- 210000003734 kidney Anatomy 0.000 description 1
- 201000011486 lichen planus Diseases 0.000 description 1
- 201000007270 liver cancer Diseases 0.000 description 1
- 208000014018 liver neoplasm Diseases 0.000 description 1
- 201000005202 lung cancer Diseases 0.000 description 1
- 208000020816 lung neoplasm Diseases 0.000 description 1
- 201000004593 malignant giant cell tumor Diseases 0.000 description 1
- 238000004949 mass spectrometry Methods 0.000 description 1
- 239000012092 media component Substances 0.000 description 1
- 201000001441 melanoma Diseases 0.000 description 1
- 210000002418 meninge Anatomy 0.000 description 1
- 239000007769 metal material Substances 0.000 description 1
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 description 1
- 238000000386 microscopy Methods 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 201000006417 multiple sclerosis Diseases 0.000 description 1
- 201000006938 muscular dystrophy Diseases 0.000 description 1
- 206010028417 myasthenia gravis Diseases 0.000 description 1
- 208000009091 myxoma Diseases 0.000 description 1
- 230000002956 necrotizing effect Effects 0.000 description 1
- 210000000653 nervous system Anatomy 0.000 description 1
- 208000004235 neutropenia Diseases 0.000 description 1
- 208000004649 neutrophil actin dysfunction Diseases 0.000 description 1
- 229920001778 nylon Polymers 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 150000002894 organic compounds Chemical class 0.000 description 1
- 201000008482 osteoarthritis Diseases 0.000 description 1
- 208000003388 osteoid osteoma Diseases 0.000 description 1
- 210000001672 ovary Anatomy 0.000 description 1
- 229960001592 paclitaxel Drugs 0.000 description 1
- 210000000496 pancreas Anatomy 0.000 description 1
- 230000010412 perfusion Effects 0.000 description 1
- 208000028169 periodontal disease Diseases 0.000 description 1
- 239000012782 phase change material Substances 0.000 description 1
- 238000009832 plasma treatment Methods 0.000 description 1
- 206010035653 pneumoconiosis Diseases 0.000 description 1
- 229920000573 polyethylene Polymers 0.000 description 1
- 238000007639 printing Methods 0.000 description 1
- 230000000069 prophylactic effect Effects 0.000 description 1
- 210000002307 prostate Anatomy 0.000 description 1
- 230000002685 pulmonary effect Effects 0.000 description 1
- ZAHRKKWIAAJSAO-UHFFFAOYSA-N rapamycin Natural products COCC(O)C(=C/C(C)C(=O)CC(OC(=O)C1CCCCN1C(=O)C(=O)C2(O)OC(CC(OC)C(=CC=CC=CC(C)CC(C)C(=O)C)C)CCC2C)C(C)CC3CCC(O)C(C3)OC)C ZAHRKKWIAAJSAO-UHFFFAOYSA-N 0.000 description 1
- 208000002574 reactive arthritis Diseases 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 206010039073 rheumatoid arthritis Diseases 0.000 description 1
- 206010039083 rhinitis Diseases 0.000 description 1
- 210000003296 saliva Anatomy 0.000 description 1
- 239000000523 sample Substances 0.000 description 1
- 201000000306 sarcoidosis Diseases 0.000 description 1
- 239000004065 semiconductor Substances 0.000 description 1
- 231100000879 sensorineural hearing loss Toxicity 0.000 description 1
- 208000023573 sensorineural hearing loss disease Diseases 0.000 description 1
- 201000001223 septic arthritis Diseases 0.000 description 1
- 238000012163 sequencing technique Methods 0.000 description 1
- 201000009890 sinusitis Diseases 0.000 description 1
- QFJCIRLUMZQUOT-HPLJOQBZSA-N sirolimus Chemical compound C1C[C@@H](O)[C@H](OC)C[C@@H]1C[C@@H](C)[C@H]1OC(=O)[C@@H]2CCCCN2C(=O)C(=O)[C@](O)(O2)[C@H](C)CC[C@H]2C[C@H](OC)/C(C)=C/C=C/C=C/[C@@H](C)C[C@@H](C)C(=O)[C@H](OC)[C@H](O)/C(C)=C/[C@@H](C)C(=O)C1 QFJCIRLUMZQUOT-HPLJOQBZSA-N 0.000 description 1
- 229960002930 sirolimus Drugs 0.000 description 1
- 201000000849 skin cancer Diseases 0.000 description 1
- 210000003625 skull Anatomy 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 210000000278 spinal cord Anatomy 0.000 description 1
- 238000004544 sputter deposition Methods 0.000 description 1
- 206010041823 squamous cell carcinoma Diseases 0.000 description 1
- 210000000130 stem cell Anatomy 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 210000002784 stomach Anatomy 0.000 description 1
- 230000002459 sustained effect Effects 0.000 description 1
- 208000011580 syndromic disease Diseases 0.000 description 1
- 201000004595 synovitis Diseases 0.000 description 1
- 201000000596 systemic lupus erythematosus Diseases 0.000 description 1
- RCINICONZNJXQF-MZXODVADSA-N taxol Chemical compound O([C@@H]1[C@@]2(C[C@@H](C(C)=C(C2(C)C)[C@H](C([C@]2(C)[C@@H](O)C[C@H]3OC[C@]3([C@H]21)OC(C)=O)=O)OC(=O)C)OC(=O)[C@H](O)[C@@H](NC(=O)C=1C=CC=CC=1)C=1C=CC=CC=1)O)C(=O)C1=CC=CC=C1 RCINICONZNJXQF-MZXODVADSA-N 0.000 description 1
- 210000001550 testis Anatomy 0.000 description 1
- 229920001169 thermoplastic Polymers 0.000 description 1
- 229920001187 thermosetting polymer Polymers 0.000 description 1
- 239000004416 thermosoftening plastic Substances 0.000 description 1
- 230000004614 tumor growth Effects 0.000 description 1
- 210000003708 urethra Anatomy 0.000 description 1
- 210000003932 urinary bladder Anatomy 0.000 description 1
- 210000002700 urine Anatomy 0.000 description 1
- 210000004291 uterus Anatomy 0.000 description 1
- 210000001215 vagina Anatomy 0.000 description 1
- 210000003905 vulva Anatomy 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M23/00—Constructional details, e.g. recesses, hinges
- C12M23/42—Integrated assemblies, e.g. cassettes or cartridges
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M23/00—Constructional details, e.g. recesses, hinges
- C12M23/34—Internal compartments or partitions
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M25/00—Means for supporting, enclosing or fixing the microorganisms, e.g. immunocoatings
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
- C12M41/12—Means for regulation, monitoring, measurement or control, e.g. flow regulation of temperature
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
- C12M41/26—Means for regulation, monitoring, measurement or control, e.g. flow regulation of pH
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
- C12M41/30—Means for regulation, monitoring, measurement or control, e.g. flow regulation of concentration
- C12M41/32—Means for regulation, monitoring, measurement or control, e.g. flow regulation of concentration of substances in solution
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
- C12M41/30—Means for regulation, monitoring, measurement or control, e.g. flow regulation of concentration
- C12M41/34—Means for regulation, monitoring, measurement or control, e.g. flow regulation of concentration of gas
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
- C12M41/30—Means for regulation, monitoring, measurement or control, e.g. flow regulation of concentration
- C12M41/36—Means for regulation, monitoring, measurement or control, e.g. flow regulation of concentration of biomass, e.g. colony counters or by turbidity measurements
Definitions
- the present invention relates to cassettes for cell culture and an analytical and preparative incubator for housing the cassettes.
- the invention relates to a cassette for culturing adherent cells comprising a first film and a second film, wherein the first and/or second film comprises at least one sensor.
- At least one film of the cassette is coated with at least one compound selected from the group consisting of a nutrient, a growth hormone, a cytokine, an antibiotic, an adhesion molecule, a mineral, an antibody, a nucleic acid probe, a primer, an oligonucleotide, an antisense oligonucleotide, a ligand, an enzyme, cell components, cell markers, cells, a dye and a test compound.
- a compound selected from the group consisting of a nutrient, a growth hormone, a cytokine, an antibiotic, an adhesion molecule, a mineral, an antibody, a nucleic acid probe, a primer, an oligonucleotide, an antisense oligonucleotide, a ligand, an enzyme, cell components, cell markers, cells, a dye and a test compound.
- the at least one sensor is present on the first and/or second film.
- the senor detects at least one of temperature, pH, cell density, O 2 , CO 2 , protein, nucleic acid, apoptotic markers, nutrient levels and the rate of consumption of a media component.
- At least a first or second film comprises an adherent layer to which cells can adhere.
- cells adhere to discrete locations on the adherent layer.
- the cassette comprises a frame.
- the at least one sensor is present on the first and/or second film and/or the frame.
- the cassette further comprises an inlet for introducing fluids and an outlet for removing fluids.
- the film comprises a material selected from the group consisting of: polymer films, polyester, polystyrene, surlyn, polyolefins, cyclic olefin polymers, poly(methyl metacrylate). PVDF membranes, cellulose membranes and nylon-semi permeable membranes.
- tissue sample is cultured in the cassette.
- the cells are removed from the layer to which they adhere by a method selected from the group consisting of: increasing or decreasing the pH, increasing or decreasing light exposure, and enzymatic degradation.
- the cassette comprises more than one compartment.
- each compartment comprises a sensor.
- the sensor of each compartment is different.
- the film comprising each compartment is coated with at least one compound selected from the group consisting of a nutrient, a growth hormone, a cytokine, an antibiotic, an adhesion molecule, a mineral, an antibody, a nucleic acid probe, a primer, an oligonucleotide, an antisense oligonucleotide, a ligand, an enzyme, cell components, cell markers, cells, a dye and a test compound.
- a compound selected from the group consisting of a nutrient, a growth hormone, a cytokine, an antibiotic, an adhesion molecule, a mineral, an antibody, a nucleic acid probe, a primer, an oligonucleotide, an antisense oligonucleotide, a ligand, an enzyme, cell components, cell markers, cells, a dye and a test compound.
- the invention also relates to a method of culturing cells comprising incubating cells in the cassette.
- the method includes a step of feeding the cells, wherein feeding is automated.
- the method includes a step of passaging the cells, wherein passaging is automated.
- At least a first or second film comprises an adherent layer to which cells can adhere.
- cells adhere to discrete locations on the adherent layer.
- the cells are removed from the layer to which they adhere by a method selected from the group consisting of: increasing or decreasing the pH, increasing or decreasing light exposure, and enzymatic degradation.
- At least one film of the cassette is coated with a compound of interest.
- the cassette is housed in an incubator.
- the invention also relates to a method of treating a disease in a subject in need thereof comprising; removing cells from the subject; transferring the cells into the cassette, culturing the cells in the presence of at least one agent that is identified as treating the disease; and reintroducing the cells into the subject, thereby treating the disease.
- the method further comprises measuring the level of an activity of the cell known to be associated with a disease and/or measuring the level of expression of a marker known to be associated with a disease: and comparing the level of activity or expression before and after the step of culturing.
- the method further comprises comparing the level or activity of cells of the subject with the level or activity of cells of a control subject that is diagnosed with the disease.
- modulation comprises a decrease in activity and/or expression.
- modulation comprises an increase in activity and/or expression.
- the agent is present in culture media.
- the first and or second film is coated with the agent.
- the disease is selected from the group recited herein below.
- the invention also relates to a method of diagnosing a subject comprising: removing cells from the subject; transferring the cells into a cassette; measuring the level of an activity of the cell known to be associated with a disease and/or measuring the level of expression of a marker known to be associated with a disease.
- the method further comprises a step of comparing the level or activity of cells of the subject with the level or activity of cells of a control subject that is not diagnosed with the disease.
- the invention also relates to a method of monitoring the progression of a disease in a subject comprising: removing cells from the subject; transferring the cells to a cassette; measuring the level of an activity of the cell known to be associated with a disease and/or measuring the level of expression of a marker known to be associated with a disease, wherein the level of an activity of a cell or the level of expression of a marker is indicative of the progression of the disease.
- the method further comprises a step of comparing the activity or expression to the activity or expression of cells isolated from the subject at an earlier time point, wherein an increase or a decrease in the level of activity or expression as compared to an earlier time point indicates the progression of the disease.
- the invention also relates to a method of designing a treatment protocol for a subject diagnosed with a disease comprising; removing cells from the subject; transferring the cells into a cassette, culturing the cells in the presence of at least one agent that is identified as increasing or decreasing the function of a cell; measuring the level of an activity of the cell known to be associated with a disease and/or measuring the level of expression of a marker known to be associated with a disease, wherein, if the agent increases or decreases the function of the cells of the subject, treating the subject with the agent.
- the agent is present in culture media.
- the agent is present on at least the first or second film.
- the invention also relates to a method of screening for a compound for treating a disease comprising: culturing cells isolated from a subject diagnosed with the disease in a cassette in the presence of a test compound; measuring the level of an activity of the cell known to be associated with a disease and/or measuring the level of expression of a marker known to be associated with a disease.
- modulation of activity or expression identifies the compound as treating the disease.
- the method further comprises a step of comparing the level of activity or expression of cells cultured in the presence of the compound with the level of activity or expression of cells cultured in the absence of the compound.
- the invention also relates to a method of screening for a compound comprising an activity of interest comprising: culturing cells in a cassette in the presence of a test compound; and; measuring the level of the activity.
- the method further comprises a step of comparing the level of activity or expression of cells cultured in the presence of the compound with the level of activity or expression of cells cultured in the absence of the compound.
- the invention also relates to a method of producing a molecule of interest comprising culturing cells engineered to produce the molecule in a cassette, under conditions wherein the molecule is produced.
- the molecule is a protein or an antibody.
- the method further comprises isolating the molecule from the cassette.
- an incubator in one embodiment, there is provided an incubator.
- the incubator includes a housing and a computing device disposed within the housing.
- the incubator also includes a chassis that is coupled to the housing and configured to receive a plurality of cassettes for culturing cells.
- the chassis includes a plurality of ports that are coupled to the computing device and configured to interface with corresponding connectors of the cassettes that receive data from sensors disposed within the cassettes.
- the incubator includes a fluidic manifold that houses a media.
- a conduit of the fluidic manifold includes a plurality of nodes configured to interface with corresponding nodes on cassettes.
- the computing device is configured to receive a schedule of processes to apply to cell cultures in cassettes; receive and store data regarding environmental conditions within the cassettes; and operate the incubator according to the schedule of processes and received data regarding the environmental conditions.
- the computing device is configured to operate the incubator according to received data regarding the temperature, gas levels, pH level, reagent concentrations, or cell density within a cassette.
- the computing device may be configured to operate the fluidic manifold to passage a cell culture in a cassette.
- the computing device may be configured to operate the fluidic manifold to flow a media into a cell culture in a cassette.
- the computing device may be configured to operate the fluidic manifold to extract contents of a cassette.
- the computing device is configured to store the received data on a remote server. In many embodiments, the computing device is configured to send an alert to a user when a condition of a process is met. The computing device may be configured to wait for a response from a user before executing a process on a cell culture in a cassette.
- the incubator includes an image capturing device coupled to the computing device.
- the computing device is configured to operate the robotic arm to position the image capturing device over a cassette.
- the incubator includes a display for displaying the received data regarding the environmental conditions within the cassettes.
- a method of operating an incubator includes storing, by a computing device, parameters for a schedule of processes to apply to cell cultures in cassettes. The method also monitors data regarding environmental conditions within the cassettes. The method includes matching received data about the environment conditions with at least one parameter for a process on the schedule. The method includes applying the process to the cell culture within the cassettes.
- the method includes receiving the parameters from a user interface of the incubator. In other embodiments, the method includes receiving the parameters from a remote server.
- the method may include monitoring data received through a port coupled to a chassis of the incubator.
- the method may include passaging the cell culture in a cassette.
- the method may include flowing a media into a cassette.
- the method may include extracting the cell culture from a cassette.
- the system will include liquid handling robotics, especially microtiter (SBS) format systems, which enable the addition and removal of samples or reagents to and from the cassettes, thereby enabling reagents such as drugs, dyes, or fluorescent probes to be added to the cells, or enabling sampled cells or media to be removed from the cassette.
- SBS microtiter
- the system will have the ability to perform analytical functions, including microscopy, mass spectrometry, and nucleic acid sequencing.
- the system will have the capability to genotype or sequence cells to confirm the identity of cell lines without the need for human intervention. This technology can further identify the presence of and nature of contaminants.
- the system will be used to propagate primary culture isolates, including cancer and stem cells, or will be used to grow sheets or masses of organ tissues, such as layers of epithelial and other cells needed for skin grafts.
- the system will be able to function in a diagnostic capacity, especially for the detection of diseases.
- microfluidic systems will be disposable to maintain sterility, and in other embodiments the system and its components will be sterilizable by internal or external sterilization methods. These include the application of chemicals such as bleach, or heat, or pressure, or sterilizing gases, or plasma treatment, or combinations thereof.
- the system may include cassettes with film battery or capacitors that enable the cassette itself to perform work, such as monitoring sensors, injecting reagents, or extracting samples.
- the multi-layered coated film on each cassette will have layers that confer a function, such as enabling or optimizing cell adhesion, serving as optical filters, or serving to minimize or maximize the transfer of fluids to each cassette.
- cassettes will be connected in bricks of 10 or more cassettes, enabling large numbers of cassettes to be easily installed or removed from the system.
- FIG. 1 presents a cassette for culturing adherent cells comprising a first and second film
- FIG. 2 presents a schematic diagram of an exemplary incubator that monitors and processes cell cultures disposed in cassettes
- FIG. 3 depicts an exemplary incubator connected over a computer network to one or more remote servers
- FIG. 4 presents the interior of an exemplary incubatory
- FIG. 5 presents an exemplary incubator
- FIG. 6 presents the interior of an exemplary table top model of an incubator
- FIG. 7 prevents an exemplary table top model of an incubator.
- the invention relates to containers or cassettes for culturing adherent cells.
- the cassettes of the various embodiments offer numerous advantages over cell culture containers known in the art.
- the cassettes and incubator permit automated detection of the cell culture environment, and automated passaging of cells, feeding of cells and sampling of cells.
- the cassettes also facilitate culture of large numbers of adherent cells in the presence of one or more compounds of interest.
- the cassettes allow for adherence of cells via methods that do not require chemical modification to detach the cells from the surface to which they adhere.
- the incubators have an increased capacity for housing cells in culture and are a designed to allow for automated monitoring and maintenance of the culture environment including but not limited to maintenance of pH, O 2 , CO 2 , and cell density.
- the incubators provide for automated sampling and imaging of cells in culture.
- the cassettes and incubator provide significant increases in the efficiency of methods of culturing adhesion cells.
- cassette means a sealed container for housing cells/and or tissues and culture media.
- a “film” comprises any material suitable for containing cells, including but not limited to polymer films, for example, polyester, polystyrene, Surlyn, polyolefins, cyclic olefin copolymers (COC), or poly(methylmethacrylate) (PMMA), membranes, for example, PVDF, cellulose, or Nylon semi-permeable membranes.
- the film is metallized to minimize gas transfer
- cells adhere to a tie layer present on the film.
- a film is coated with at least one compound including but not limited to a nutrient, a growth factor, a hormone, a cytokine, an antibiotic, an adhesion molecule, a mineral, an antibody, a nucleic acid, including but not limited to nucleic acid probes, primers and oligonucleotides, ligands, enzymes such as polymerases, proteases, isomerases, cell components or cell markers, small molecules and dyes.
- the compound is living or dead cells (e.g., for use as a viral target, or a baseline control for determining cell concentration or baseline or basal levels of target protein expression).
- the cells can be stained or treated to allow for detection.
- the “film” is coated with a test compound to be screened for a particular activity of interest.
- the film can be of any dimensions such that a surface for culturing the desired number of cells is provided.
- the thickness of the film can be such that diffusion of material across the surface of the film can occur. In one embodiment, the thickness of the film is from 1 ⁇ M to 1 mM, for example, 10 ⁇ M to 500 ⁇ M, or 20 ⁇ M to 100 ⁇ M.
- the film comprises a tie layer(s) comprising a material that can be separated from or peeled off of the film.
- a tie layer comprises a photo-depolymerizing material, or a material that, for example material that is thermoreversible, chemoreversible or enzymatically cleavable. Under the appropriate condition (light, heat, chemical environment) the tie layer reverts to monomers or small polymers that dissolve and release the cells.
- film comprises a tie layer composed of a phase change material that allows the tie layer to be peeled off the film without reverting to monomers and small polymers that eventually dissolve, thereby providing a sheet of cells.
- a “frame” refers to a structure that is used in association with the film comprising a cassette.
- a “frame” can be either solid or flexible.
- a “frame” comprises at least one sensor as defined herein.
- a “frame” is located either between a first and second film or on the exterior of a certain layer of film, for example, around the perimeter of the film.
- the frame comprises polystyrene, PMMA, polyolefins such as polyethylene, surlyn, engineering thermoplastics and any thermoplastic, thermoset or composite thereof.
- the film in combination with the frame is under tension.
- the cassette can include frangible sealed regions or compartments containing, for example, a liquid or solid where a sustained squeeze just prior to use will burst the seal and swirl the components together.
- frangible sealed regions or compartments containing, for example, a liquid or solid where a sustained squeeze just prior to use will burst the seal and swirl the components together.
- the multiple component pouches with internal frangible seals afford the opportunity to conveniently and inexpensively deliver portions with time-of-use mixing. This allows for extending the shelf life of cassettes dramatically. All of the biologically active components are maintained in a fluid chamber that the user or machine can burst in.
- the frangible film is Surlyn, which does not contribute particulars to the mixtures when the seal is broken.
- a “sensor” detects or measures one or more properties of the cell and/or environment inside the cassette.
- a “sensor” can monitor, modify, record, indicate, or otherwise respond to one or more properties of the cell and/or the environment inside a cassette.
- a sensor detects at least one of temperature, pH, O 2 , CO 2 and cell density.
- a sensor detects protein expression by detecting fluorescence, for example by a FISH assays.
- the sensor is ligand-based or ligand-specific for detection of, for example, specific proteins or nucleic acids, apoptotic markers, nutrient levels, and/or the consumption rates of various media constituents.
- a sensor can be located on the film and/or the frame comprising the cassette.
- a “film” is coated with a compound useful for altering a property of a cell or tissue and/or supporting cell growth and viability and/or facilitating adhesion of the cells to the film.
- a “compound” includes but is not limited to a nutrient, a growth factor, a hormone, a cytokine, an antibiotic, an adhesion molecule, a mineral, an antibody, a nucleic acid, including a nucleic acid probe, an antisense molecule, a primer and an oligonucleotide, a ligand for a receptor of interest, enzymes, for example, including but not limited to polymerases, proteases, isomerases etc. . . .
- the compound can be a cell(s), either living or dead, for use as a viral target, for defining a baseline level of cellular activity and/or protein or mRNA expression; and to establish a baseline for cell concentration.
- the cells can be living or dead, and stained or otherwise treated to facilitate detection.
- the “film” can also be coated with a test compound to be screened for a particular activity of interest or a therapeutic agent.
- the cassette comprises a first and second layer of film, and further comprises one or more compartments.
- a “compartment” refers to an enclosed section of the cassette, that can be used to house cells of interest such that they are separated from and are not in contact with cells that are present in the cassette but are located outside of the compartment.
- a “compartment” can include a sensor.
- the regions of the film comprising a compartment may be coated with at least one compound of interest.
- the compartment can comprise a particular cell media that is either different from or identical to media present in other compartments of the cassette.
- fluid is introduced to or removed from a “compartment” via a unique fluid line.
- the invention also provides for cassettes wherein replicate samples of cells are adhered to the film at discrete locations but are not confined to a “compartment” as defined herein.
- modulate or “modulation” refers to increase or decrease.
- “decrease” as it refers to the level of activity of a cell or the level of expression of a marker, for example a protein, mRNA or antibody of interest by a cell means that activity or expression is 1, 2, 3, 4, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 100, 500, 1000 or 10,000-fold less before or after administration to or incubation in the presence of a cell with a compound of interest.
- “decrease” as it refers to the level of activity of a cell or the level of expression of a marker, for example a protein, mRNA or antibody of interest by a cell means that activity or expression is 1, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 99 or 100% before or after administration to or incubation in the presence of a cell with a compound of interest.
- a marker for example a protein, mRNA or antibody of interest by a cell
- activity or expression is 1, 2, 3, 4, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 100, 500, 1000 or 10,000-fold or more before or after administration to or incubation in the presence of a cell with a compound of interest.
- mRNA or antibody of interest by a cell is 1, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 99 or 100% greater before or after administration to or incubation in the presence of a cell with a compound of interest.
- the term “disease” includes any one or more of the following autoimmune diseases or disorders: diabetes mellitus, arthritis (including rheumatoid arthritis, juvenile rheumatoid arthritis, osteoarthritis, psoriatic arthritis), multiple sclerosis, myasthenia gravis, systemic lupus erythematosis, autoimmune thyroiditis, dermatitis (including atopic dermatitis and eczematous dermatitis), psoriasis, Sjögren's Syndrome, including keratoconjunctivitis sicca secondary to Sjögren's Syndrome, alopecia areata, allergic responses due to arthropod bite reactions, Crohn's disease, aphthous ulcer, ulceris, conjunctivitis, keratoconjunctivitis, ulcerative colitis, asthma, allergic asthma, cutaneous lupus erythematosus, scleroderma, vaginitis,
- disease refers to any one of Wilson's disease, spinocerebellar ataxia, prion disease, Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis, amyloidosis, Alzheimer's disease, Alexander's disease, alcoholic liver disease, cystic fibrosis, Pick's Disease, spinal muscular dystrophy or Lewy body dementia.
- Disease also includes any one of rheumatoid spondylitis; post ischemic perfusion injury: inflammatory bowel disease; chronic inflammatory pulmonary disease, eczema, asthma, ischemia/reperfusion injury, acute respiratory distress syndrome, infectious arthritis, progressive chronic arthritis, deforming arthritis, traumatic arthritis, gouty arthritis, Reiter's syndrome, acute synovitis and spondylitis, glomerulonephritis, hemolytic anemia, aplastic anemia, neutropenia, host versus graft disease, allograft rejection, chronic thyroiditis.
- Graves' disease primary binary cirrhosis, contact dermatitis, skin sunburns, chronic renal insufficiency, Guillain-Barre syndrome, uveitis, otitis media, periodontal disease, pulmonary interstitial fibrosis, bronchitis, rhinitis, sinusitis, pneumoconiosis, pulmonary insufficiency syndrome, pulmonary emphysema, pulmonary fibrosis, silicosis, or chronic inflammatory pulmonary disease.
- Disease also refers to any one of cancer, tumor growth, cancer of the colon, breast, bone, brain and others (e.g., osteosarcoma, neuroblastoma, colon adenocarcinoma), chronic myelogenous leukemia (CML), acute myeloid leukemia (AML), acute promyelocytic leukemia (APL), cardiac cancer (e.g., sarcoma, myxoma, rhabdomyoma, fibroma, lipoma and teratoma); lung cancer (e.g., bronchogenic carcinoma, alveolar carcinoma, bronchial adenoma, sarcoma, lymphoma, chondromatous hamartoma, mesothelioma): various gastrointestinal cancer (e.g., cancers of esophagus, stomach, pancreas, small bowel, and large bowel); genitourinary tract cancer (e.g., kidney, bladder and
- Ewing's sarcoma malignant lymphoma, multiple myeloma, malignant giant cell tumor chordoma, osteochronfroma, benign chondroma, chondroblastoma, chondromyxofibroma, osteoid osteoma and giant cell tumors); cancers of the nervous system (e.g., of the skull, meninges, brain, and spinal cord); gynecological cancers (e.g., uterus, cervix, ovaries, vulva, vagina); hematologic cancer (e.g., cancers relating to blood, Hodgkin's disease, non-Hodgkin's lymphoma); skin cancer (e.g., malignant melanoma, basal cell carcinoma, squamous cell carcinoma.
- gynecological cancers e.g., uterus, cervix, ovaries, vulva, vagina
- hematologic cancer e.g
- Karposi's sarcoma moles dysplastic nevi, lipoma, angioma, dermatofibroma, keloids, psoriasis); and cancers of the adrenal glands (e.g., neuroblastoma).
- diagnosis or “identifying a patient or subject having” or “a patient or subject in need of treatment” refers to a process of determining if an individual is afflicted with a disease or ailment.
- a subject is said to be treated for a disease, if following administration of an agent, one or more symptoms of the disease are decreased or eliminated.
- Treatment is defined as the application or administration of a compound identified as treating a disease of interest, to a subject or patient, or application or administration of a compound identified as treating a disease of interest to an isolated tissue or cell line from a subject or patient, who has a disease or disorder, with the purpose to cure, heal, alleviate, relieve, alter, remedy, ameliorate, improve or affect the disease or disorder, or symptoms of the disease or disorder.
- treatment or “treating” is also used herein in the context of administering agents prophylactically.
- effective dose or “effective amount” or “effective dosage” or “therapeutic dosage” is defined as an amount sufficient to achieve or at least partially achieve the desired effect.
- therapeutically effective dose” and “therapeutically effective amount” are defined as an amount sufficient to cure or at least partially arrest the disease and its complications in a patient already suffering from the disease.
- treating refers to preventing the onset of disease and/or reducing, delaying, or eliminating disease symptoms, such as an increase in the extracellular level of a group of cytokines.
- treating is meant restoring the patient or subject to the basal state as defined herein, and/or to prevent a disease in a subject at risk thereof.
- treating means arresting or otherwise ameliorating symptoms of a disease.
- patient or “subject” refers to a mammal that is diagnosed with a disease associated with an increase in the extracellular level of a group of cytokines.
- patient or “subject” includes human and other mammalian subjects that receive either prophylactic or therapeutic treatment.
- control subject means a subject that has not been diagnosed with a disease and/or does not exhibit any detectable symptoms associated with the disease.
- mammal refers to any mammal including but not limited to human, mouse, rat, sheep, monkey, goat, rabbit, hamster, horse, cow or pig.
- non-human mammal refers to any mammal that is not a human.
- basic state refers to the extracellular level of a group of cytokines of an individual who is not susceptible to a disease and who has no symptoms of a disease.
- monitoring the treatment means determining whether, following treatment of a subject, for example, a subject diagnosed with a disease associated with an increased extracellular level of a group of cytokines, the subject has been treated such that the symptoms of the disease are arrested or otherwise ameliorated and/or the disease and/or its attendant symptoms are alleviated or abated.
- pharmaceutically acceptable carrier refers to a carrier for the administration of a therapeutic agent.
- exemplary carriers include saline, buffered saline, dextrose, water, glycerol, ethanol, and combinations thereof.
- small molecule refers to a non-peptidic, non-oligomeric organic compound either synthesized in the laboratory or found in nature.
- Small molecules can refer to compounds that are “natural product-like”.
- the term “small molecule” is not limited to “natural product-like” compounds. Rather, a small molecule is typically characterized in that it contains several carbon-carbon bonds, and has a molecular weight of less than 1500, although this characterization is not intended to be limiting for the purposes of the present invention. Examples of “small molecules” that occur in nature include, but are not limited to, taxol, dynemicin, and rapamycin. In certain other preferred embodiments, natural-product-like small molecules are utilized.
- the cassettes and incubator can be used to culture adherent cells under conditions that are precisely regulated.
- the cells can be maintained in culture by automated methods.
- the cassettes and incubator are useful for performing methods of treating a disease, methods of diagnosing a disease, methods of monitoring disease treatment and methods of designing a treatment protocol.
- the cassettes and incubator facilitate screening for a compound, for example, a therapeutic.
- FIG. 1 presents one embodiment of a cassette comprising a first and second layer of film and a frame.
- the cassette can be prepared by a method including preparing a frame that is, for example, injected molded, comprises a laminate-layered core or cast core or center core, is 3D printed, machined or lasercut.
- the frame is surrounded on the top and bottom by film, membrane, multilayer coated film or a combination thereof.
- the frame is surround on the top and bottom with a thick film that is die-stamped, laser cut, etc. . . .
- the film is coated at least by any of the following methods: roll to roll coating, printing of certain regions of the film, for example PRP islets to which discrete populations of cells will adhere, reagents used for detection, spots, lanes or other structures, hydrophobic and hydrophilic regions to channel liquid flow within the cassette.
- the file is coated with a metallic substance or other vapor or gas control coatings, for example by chemical vapor deposit (CVD) or sputtering, gravure coating, skiving and any other method of applying a coating reagent, ink or dye to a film or surface known in the art.
- CVD chemical vapor deposit
- sputtering gravure coating
- skiving any other method of applying a coating reagent, ink or dye to a film or surface known in the art.
- cells are introduced into a cassette using a sterilizable and/or disposable, microfluidic manifold located in the rear of the chamber that controls the flow of median to each of the cassettes.
- a micro plate is inserted into the machine, and the cells are individually aspirated from the plate via a disposable or reusable sterilizable capillary device that interfaces with the manifold.
- the capillary is robotically moved or and through gates and microfluidics the flow can be directed.
- a robot that traverses the entire back of the system, has a capillary injection port that butts against the array or the cassettes themselves and injects the cells. Movement of media in an out the cassette can occur via this process.
- the user can lift a corner of the film and inject via pipet (such a liftable corner is depicted in FIG. 1 ) then reseal (for example, via an adhesive); or the user can inject through a re-sealable port that is built into the film or frame or both of the cassette.
- FIG. 2 represents a schematic diagram of an exemplary incubator 200 that monitors and processes cell cultures disposed in cassettes.
- the incubator 200 includes a programmable computing device 205 that controls the incubator's operations.
- the computing device 205 includes a user interface, a processor, and a memory.
- the memory stores instructions for operating the incubator 200 .
- the computing device 205 receives parameters from a user for processing cell cultures.
- the computing device 205 executes instructions according to the parameters to control components of the incubator 200 .
- the computing device 205 is configured to communicate over computer networks 305 with remote servers 310 .
- the remote servers 310 may store data from the computing device 205 or communicate with users according to instructions from the computing device 205 , as described in more detail below.
- the incubator includes a chassis 210 disposed in a housing 211 .
- the chassis 210 includes shelves that receive and support cassettes containing cell cultures.
- the shelves have a plurality of ports 215 a , 215 b , . . . (“ 215 ”).
- Each shelf has a port 215 that is coupled to the computing device 205 .
- the shelf's port 215 interfaces with a connector on the cassette that is coupled to the cassette's circuitry, which receives data from sensors disposed within the cassette.
- the port 215 includes one or more connections for supplying power to the cassette's circuitry and/or receiving data collected by the circuitry's sensors. Data received through this port 215 is transmitted to the computing device 205 .
- the incubator 200 includes a fluidic manifold 220 .
- a fluidic manifold 220 has a chamber 225 that houses a media of interest useful for cell culture.
- the chamber 225 receives cell cultures extracted from cassettes.
- a conduit 230 extends from the chamber 225 to shelves in the chassis 210 .
- the conduit 230 includes nodes 235 a , 235 b , . . . (“ 235 ”) that can be coupled to corresponding nodes on cassettes.
- the figure depicts a single conduit that connects to each of the cassettes alternative embodiments may include multiple conduits, each of which is coupled to a distinct cassette or a subset of the cassettes.
- a pump 240 coupled to the fluidic manifold 220 can propel media in the chamber 225 through a conduit into one or more of the cassettes.
- a vacuum 245 coupled to the fluidic manifold 220 extracts the contents of one or more cassettes into a chamber.
- the fluidic manifold 220 extracts the contents of a cassette into the same chamber used for propelling media into cassettes in the incubator.
- the fluidic manifold 220 includes separate chambers for housing media to propel into cassettes and receiving contents extracted from cassettes.
- the incubator 200 has one or more image capturing devices and robotic arms for positioning the devices within the housing 211 .
- the computing device 205 operates the robotic arms to position an image capturing device over a cell culture in a cassette.
- the computing device 205 operates the image capturing device to obtain an image of a cell culture.
- FIG. 3 depicts the incubator 200 connected over a computer network 305 to one or more remote servers 310 .
- the computing device 205 connects to the remote server(s) 310 via a local area connection (LAN), wide area connection (WAN), the Internet, or any computer network.
- the computing device 205 transmits data regarding a cell culture, for example, pH and/or cell density, and environmental conditions within cassettes to a remote server 310 for storage.
- the server 315 stores the data.
- the computing device 205 transmits messages to a remote server 315 for communicating with a user of the incubator.
- the remote server 315 sends the messages to a user.
- a user introduces cells in combination with a media of interest into the cassettes and inserts the cassettes into the shelves of the incubator 200 .
- the computing device 205 detects the presence of the cassette.
- the computing device 205 powers the cassette's circuitry by supplying a power signal through the port 215 .
- the computing device 205 initializes a file for storing data on the environmental conditions of the cassette.
- the computing device 205 receives data collected by sensors in the cassette, such as temperature, cell density, and pH.
- the computing device 205 stores the data in the file associated with the cassette. In other embodiments, the computing device 205 discards the data until the device 205 receives processing instructions from the user.
- a user programs the computing device 205 to process cell cultures disposed in cassettes in the incubator 200 .
- the user programs the computing device 205 from a user interface disposed on the incubator 200 .
- the user interface displays a menu of parameters for the user to select. For example, the user can select the temperature and humidity at which the incubator will maintain the environment in the cassettes. The user can select the media to introduce into the cassettes during processing.
- the user interface displays the % concentration of a gas, for example, O2 or CO2, the temperature inside the incubator, the composition and/or volume and/or pH of the media, or the amount of other substances present in the incubator's chambers, and the user can supply the incubator with additional resources, if necessary.
- the user can create a schedule of actions with respect to the cell cultures. Some of the actions may be directed to monitoring the cell cultures. For example, the user may configure the incubator to begin beeping every 6 hours, thereby alerting the user to check conditions of the cell cultures. In another example, the user may configure the incubator to send the user a short message service (SMS) communication every 6 hours, for the same purpose.
- SMS short message service
- the user may input a mobile telephone number at which the user is capable of receiving SMS communication. Every six hours, the computing device 205 sends an instruction to the remote server 310 to send a reminder message to the user s mobile telephone number. In response, the remote server 310 creates the corresponding SMS communication and sends the same to the telephone number.
- SMS short message service
- the user may configure the incubator to capture an image of the cells present in a particular cassette and e-mail the image to the user every 6 hours.
- the user may input an e-mail address.
- the computing device 205 operates a robotic arm to position an image capturing device over the cassette of interest, capture an image, and send the image and e-mail address to a remote server 310 .
- the remote server 310 creates an e-mail message with the image and sends the same to the user.
- Some of the actions may be directed to maintaining environmental conditions within the cassette.
- the user can select a desired pH level for a cell culture, an acceptable level of deviation from the desired pH level, and media for adjusting the pH level in a cassette.
- the fluidic manifold 220 flows additional media of the user's choosing into the cassette.
- the fluidic manifold 220 flows a different media selected by the user if the pH level becomes too acidic.
- the user can select a desired level of feed for the cell culture.
- the user can select a desired concentration, or a minimum concentration, of feed present in the cell culture's environment. When the concentration of feed drops below the desired or minimum level, the fluidic manifold 220 flows additional feed into the cassette.
- Some of the actions may be directed to processing a cell culture within a cassette.
- An action may depend upon elapses of time, conditions regarding the cell culture itself or the environment in the cassette (e.g., pH, temperature, cell density), or user initiative.
- the user may schedule the incubator to flow a media into ten cassettes comprising cell cultures, heat the cassettes until they reach a desired temperature, and maintain the desired temperature for a particular length of time.
- the user may schedule the incubator to monitor the pH levels of the cassettes and flow a selected media through the cassettes once the pH levels reach a user selected threshold.
- the user may schedule the incubator to monitor the cell density of cultures and passage the same once the cell density reaches a desired threshold.
- the fluidic manifold 220 flows the passaging media into the cassettes, and the vacuum extracts the cells into a chamber for retrieval by the user.
- the user may schedule the incubator to send alerts whenever a programmed condition is met. For example, a user may schedule the incubator to flow a media into cassettes when the pH level in the cassette drops below a threshold level.
- the computing device 205 continues to receive data about the pH levels from the cassettes, and when the pH levels drop to the threshold, the computing device 205 sends the user an alert (e.g., an e-mail message, an SMS message).
- an alert e.g., an e-mail message, an SMS message.
- the user may schedule the incubator to monitor the cell density and send the user an image of a culture when the detected cell density reaches a desired threshold.
- the incubator may require user approval to passage the cells, or to enact another process. If the incubator does not receive such approval, the incubator may maintain the environmental conditions of the cassette and continue sending images to the user, according to a predetermined schedule. Once the user approves an image of the cell cultures, based on its density, the incubator may passage the cell culture, or perform some other desired process.
- Processing actions may include any manipulation of the cell cultures, according to any conditions.
- the user can schedule a series of processes, based on any of the parameters described herein. For example, the user can instruct the incubator to flow medium A into the cassettes and allow the cells to incubate for a particular period of time. After the four hours have elapsed, the incubator may flow medium B into the cassettes until the pH level for each cassette has dropped to a user selected level. Then, the incubator may raise the temperature of the incubator to a desired temperature. Once the temperature reaches the desired temperature, the fluidic manifold may flow medium C into the cassettes to feed the cell cultures. The incubator may monitor the cell density of each cassette.
- the fluidic manifold may flow a passaging reagent into the cassettes, the cells are incubated for a predetermined period of time in the presence of the reagent such that the cells dissociate from the film comprising the cassette, and vacuum the cassettes' contents into a chamber from which the cells can be isolated. Processes according to similar parameters, or sequences thereof, may also be deployed.
- the user can program the computing device 205 remotely.
- the user can log into a remote computing device 205 that executes an application that instructs the sequence of events required to perform a particular experimental analysis.
- the application may include a similar user interface to the one depicted on the incubator.
- the user may create a schedule of actions for the incubator, as described herein.
- the computing device 205 executing the application transmits the user controls over a computer network 305 to the incubator, and the incubator's computing device 205 stores the controls in its memory.
- the computing device 205 receives data about environmental conditions in the cassettes through the ports 215 and store the data in files corresponding to the cassettes.
- the computing device 205 stores the data in its local memory.
- the user can access the data through the user interface. From the menu, the user can select the file corresponding to a cassette of interest, and the computing device 205 displays the cassette's data. The user can scroll through the data to view data at different intervals of time.
- the computing device 205 transmits the data over a computer network 305 to a remote server for storage. Users can access the data on the remote servers 310 . In some embodiments, using an application executing on a computing device, a user can log into an account on the remote server 310 . The user may view the data collected by the incubator 200 and transmitted to the remote server 310 . In this manner, the user may monitor the progress of the cell cultures and the environmental conditions through which the cell cultures have progressed.
- embodiments may be implemented at least in part in any conventional computer programming language. For example, some embodiments may be implemented in a procedural programming language (e.g., “C”), or in an object oriented programming language (e.g., “C++”). Other embodiments may be implemented as preprogrammed hardware elements (e.g., application specific integrated circuits, FPGAs. and digital signal processors), or other related components.
- C procedural programming language
- object oriented programming language e.g., “C++”
- preprogrammed hardware elements e.g., application specific integrated circuits, FPGAs. and digital signal processors
- the disclosed apparatus and methods may be implemented as a computer program product for use with a computer system.
- Such implementation may include a series of computer instructions fixed either on a tangible medium, such as a computer readable medium (e.g., a diskette, CD-ROM. ROM, or fixed disk).
- the series of computer instructions can embody all or part of the functionality previously described herein with respect to the system.
- Such computer instructions can be written in a number of programming languages for use with many computer architectures or operating systems.
- such instructions may be stored in any memory device, such as semiconductor, magnetic, optical or other memory devices, and may be transmitted using any communications technology, such as optical, infrared, microwave, or other transmission technologies.
- such a computer program product may be distributed as a tangible removable medium with accompanying printed or electronic documentation (e.g., shrink wrapped software), preloaded with a computer system (e.g., on system ROM or fixed disk), or distributed from a server or electronic bulletin board over the network (e.g., the Internet or World Wide Web).
- a computer system e.g., on system ROM or fixed disk
- a server or electronic bulletin board over the network
- some embodiments may be implemented as a combination of both software (e.g., a computer program product) and hardware.
- Still other embodiments are implemented as entirely hardware, or entirely software.
- a cell of interest is introduced into a cassette of the invention via a sterilizable and/or disposable microfluidic manifold located in the rear of the chamber that controls the follow of medium to a cassette of the invention.
- a cell is introduced either manually or via automated methods.
- a cell is introduced in combination with a media of interest into a cassette and the cassette is inserted into an incubator according to the invention.
- the cell is cultured in the presence of at least one agent known to have a desired effect on the cell, for example, increase or decrease an activity of the cell, or increase or decrease expression of a marker of the cell.
- the cell can also be cultured in the presence of an agent identified as treating a disease. Cells are cultured at a density know to be appropriate for optimal growth of that particular cell type.
- Cells are removed from a subject in need of treatment.
- Cells that are removed can be present in a biological sample including but not limited to, blood, saliva, urine and tissue.
- the cells are transferred into the cassette of the invention and cultured in the presence of at least one agent identified as treating the disease. Following the culture period, the cells are reintroduced into the subject, for example, by injection, thereby treating the disease.
Abstract
The invention relates to cassettes for culturing cells. The invention also relates to analytical and preparative incubators for housing the cassettes. The invention also relates to methods of using the cassettes, for example, to culture cells wherein the processes of feeding and passaging cells are automated.
Description
- This application is a continuation of U.S. Nonprovisional patent application Ser. No. 16/694,285, filed Nov. 25, 2019, pending, which is a division of U.S. Nonprovisional patent application Ser. No. 15/427,887, filed Feb. 8, 2017, now U.S. Pat. No. 10,519,411, which, is a division of U.S. Nonprovisional patent application Ser. No. 14/211,711, filed Mar. 14, 2014, now U.S. Pat. No. 9,597,355, which claims priority of U.S. Provisional Patent Application No. 61/802,976, filed Mar. 18, 2013, and of U.S. Provisional Patent Application No. 61/788,745, filed Mar. 15, 2013, the entire contents of which patent applications are hereby incorporated herein by reference.
- The present invention relates to cassettes for cell culture and an analytical and preparative incubator for housing the cassettes.
- There is a need in the art for efficient and automated methods of culturing cells, especially adherent cells, that allow for culturing large numbers of cells in a single vessel.
- The invention relates to a cassette for culturing adherent cells comprising a first film and a second film, wherein the first and/or second film comprises at least one sensor.
- In one embodiment, at least one film of the cassette is coated with at least one compound selected from the group consisting of a nutrient, a growth hormone, a cytokine, an antibiotic, an adhesion molecule, a mineral, an antibody, a nucleic acid probe, a primer, an oligonucleotide, an antisense oligonucleotide, a ligand, an enzyme, cell components, cell markers, cells, a dye and a test compound.
- In another embodiment, the at least one sensor is present on the first and/or second film.
- In another embodiment, the sensor detects at least one of temperature, pH, cell density, O2, CO2, protein, nucleic acid, apoptotic markers, nutrient levels and the rate of consumption of a media component.
- In another embodiment, at least a first or second film comprises an adherent layer to which cells can adhere.
- In another embodiment, cells adhere to discrete locations on the adherent layer.
- In another embodiment, the cassette comprises a frame.
- In another embodiment, the at least one sensor is present on the first and/or second film and/or the frame.
- In another embodiment, the cassette further comprises an inlet for introducing fluids and an outlet for removing fluids.
- In another embodiment, the film comprises a material selected from the group consisting of: polymer films, polyester, polystyrene, surlyn, polyolefins, cyclic olefin polymers, poly(methyl metacrylate). PVDF membranes, cellulose membranes and nylon-semi permeable membranes.
- In another embodiment a tissue sample is cultured in the cassette. In another embodiment, the cells are removed from the layer to which they adhere by a method selected from the group consisting of: increasing or decreasing the pH, increasing or decreasing light exposure, and enzymatic degradation.
- In another embodiment, the cassette comprises more than one compartment.
- In another embodiment, each compartment comprises a sensor.
- In another embodiment, the sensor of each compartment is different.
- In another embodiment, the film comprising each compartment is coated with at least one compound selected from the group consisting of a nutrient, a growth hormone, a cytokine, an antibiotic, an adhesion molecule, a mineral, an antibody, a nucleic acid probe, a primer, an oligonucleotide, an antisense oligonucleotide, a ligand, an enzyme, cell components, cell markers, cells, a dye and a test compound.
- The invention also relates to a method of culturing cells comprising incubating cells in the cassette.
- In one embodiment, the method includes a step of feeding the cells, wherein feeding is automated.
- In another embodiment, the method includes a step of passaging the cells, wherein passaging is automated.
- In another embodiment, at least a first or second film comprises an adherent layer to which cells can adhere.
- In another embodiment, cells adhere to discrete locations on the adherent layer.
- In another embodiment, the cells are removed from the layer to which they adhere by a method selected from the group consisting of: increasing or decreasing the pH, increasing or decreasing light exposure, and enzymatic degradation.
- In another embodiment, at least one film of the cassette is coated with a compound of interest.
- In another embodiment, the cassette is housed in an incubator.
- The invention also relates to a method of treating a disease in a subject in need thereof comprising; removing cells from the subject; transferring the cells into the cassette, culturing the cells in the presence of at least one agent that is identified as treating the disease; and reintroducing the cells into the subject, thereby treating the disease.
- In one embodiment, the method further comprises measuring the level of an activity of the cell known to be associated with a disease and/or measuring the level of expression of a marker known to be associated with a disease: and comparing the level of activity or expression before and after the step of culturing.
- In another embodiment, the method further comprises comparing the level or activity of cells of the subject with the level or activity of cells of a control subject that is diagnosed with the disease.
- In another embodiment, following the step of culturing there is a modulation in activity or expression, thereby treating the disease.
- In another embodiment, modulation comprises a decrease in activity and/or expression.
- In another embodiment, modulation comprises an increase in activity and/or expression.
- In another embodiment, the agent is present in culture media.
- In another embodiment, the first and or second film is coated with the agent.
- In another embodiment, the disease is selected from the group recited herein below.
- The invention also relates to a method of diagnosing a subject comprising: removing cells from the subject; transferring the cells into a cassette; measuring the level of an activity of the cell known to be associated with a disease and/or measuring the level of expression of a marker known to be associated with a disease.
- In one embodiment, the method further comprises a step of comparing the level or activity of cells of the subject with the level or activity of cells of a control subject that is not diagnosed with the disease.
- The invention also relates to a method of monitoring the progression of a disease in a subject comprising: removing cells from the subject; transferring the cells to a cassette; measuring the level of an activity of the cell known to be associated with a disease and/or measuring the level of expression of a marker known to be associated with a disease, wherein the level of an activity of a cell or the level of expression of a marker is indicative of the progression of the disease.
- In one embodiment, the method further comprises a step of comparing the activity or expression to the activity or expression of cells isolated from the subject at an earlier time point, wherein an increase or a decrease in the level of activity or expression as compared to an earlier time point indicates the progression of the disease.
- The invention also relates to a method of designing a treatment protocol for a subject diagnosed with a disease comprising; removing cells from the subject; transferring the cells into a cassette, culturing the cells in the presence of at least one agent that is identified as increasing or decreasing the function of a cell; measuring the level of an activity of the cell known to be associated with a disease and/or measuring the level of expression of a marker known to be associated with a disease, wherein, if the agent increases or decreases the function of the cells of the subject, treating the subject with the agent.
- In another embodiment, the agent is present in culture media.
- In another embodiment, the agent is present on at least the first or second film.
- The invention also relates to a method of screening for a compound for treating a disease comprising: culturing cells isolated from a subject diagnosed with the disease in a cassette in the presence of a test compound; measuring the level of an activity of the cell known to be associated with a disease and/or measuring the level of expression of a marker known to be associated with a disease.
- In one embodiment, modulation of activity or expression identifies the compound as treating the disease.
- In another embodiment, the method further comprises a step of comparing the level of activity or expression of cells cultured in the presence of the compound with the level of activity or expression of cells cultured in the absence of the compound.
- The invention also relates to a method of screening for a compound comprising an activity of interest comprising: culturing cells in a cassette in the presence of a test compound; and; measuring the level of the activity.
- In another embodiment, the method further comprises a step of comparing the level of activity or expression of cells cultured in the presence of the compound with the level of activity or expression of cells cultured in the absence of the compound.
- The invention also relates to a method of producing a molecule of interest comprising culturing cells engineered to produce the molecule in a cassette, under conditions wherein the molecule is produced.
- In one embodiment, the molecule is a protein or an antibody.
- In another embodiment, the method further comprises isolating the molecule from the cassette.
- In one embodiment, there is provided an incubator. The incubator includes a housing and a computing device disposed within the housing. The incubator also includes a chassis that is coupled to the housing and configured to receive a plurality of cassettes for culturing cells. The chassis includes a plurality of ports that are coupled to the computing device and configured to interface with corresponding connectors of the cassettes that receive data from sensors disposed within the cassettes. The incubator includes a fluidic manifold that houses a media. A conduit of the fluidic manifold includes a plurality of nodes configured to interface with corresponding nodes on cassettes. The computing device is configured to receive a schedule of processes to apply to cell cultures in cassettes; receive and store data regarding environmental conditions within the cassettes; and operate the incubator according to the schedule of processes and received data regarding the environmental conditions.
- In some embodiments, the computing device is configured to operate the incubator according to received data regarding the temperature, gas levels, pH level, reagent concentrations, or cell density within a cassette. The computing device may be configured to operate the fluidic manifold to passage a cell culture in a cassette. The computing device may be configured to operate the fluidic manifold to flow a media into a cell culture in a cassette. The computing device may be configured to operate the fluidic manifold to extract contents of a cassette.
- In some embodiments, the computing device is configured to store the received data on a remote server. In many embodiments, the computing device is configured to send an alert to a user when a condition of a process is met. The computing device may be configured to wait for a response from a user before executing a process on a cell culture in a cassette.
- In many embodiments, the incubator includes an image capturing device coupled to the computing device. The computing device is configured to operate the robotic arm to position the image capturing device over a cassette. In some embodiments, the incubator includes a display for displaying the received data regarding the environmental conditions within the cassettes.
- In another embodiment, a method of operating an incubator includes storing, by a computing device, parameters for a schedule of processes to apply to cell cultures in cassettes. The method also monitors data regarding environmental conditions within the cassettes. The method includes matching received data about the environment conditions with at least one parameter for a process on the schedule. The method includes applying the process to the cell culture within the cassettes.
- In some embodiment, the method includes receiving the parameters from a user interface of the incubator. In other embodiments, the method includes receiving the parameters from a remote server. The method may include monitoring data received through a port coupled to a chassis of the incubator. The method may include passaging the cell culture in a cassette. The method may include flowing a media into a cassette. The method may include extracting the cell culture from a cassette.
- In some embodiments, the system will include liquid handling robotics, especially microtiter (SBS) format systems, which enable the addition and removal of samples or reagents to and from the cassettes, thereby enabling reagents such as drugs, dyes, or fluorescent probes to be added to the cells, or enabling sampled cells or media to be removed from the cassette.
- In some embodiments, the system will have the ability to perform analytical functions, including microscopy, mass spectrometry, and nucleic acid sequencing.
- In some embodiments, the system will have the capability to genotype or sequence cells to confirm the identity of cell lines without the need for human intervention. This technology can further identify the presence of and nature of contaminants.
- In some embodiments, the system will be used to propagate primary culture isolates, including cancer and stem cells, or will be used to grow sheets or masses of organ tissues, such as layers of epithelial and other cells needed for skin grafts.
- In some embodiments, the system will be able to function in a diagnostic capacity, especially for the detection of diseases.
- In some embodiments the microfluidic systems will be disposable to maintain sterility, and in other embodiments the system and its components will be sterilizable by internal or external sterilization methods. These include the application of chemicals such as bleach, or heat, or pressure, or sterilizing gases, or plasma treatment, or combinations thereof.
- In some embodiments the system may include cassettes with film battery or capacitors that enable the cassette itself to perform work, such as monitoring sensors, injecting reagents, or extracting samples.
- In some embodiments, the multi-layered coated film on each cassette will have layers that confer a function, such as enabling or optimizing cell adhesion, serving as optical filters, or serving to minimize or maximize the transfer of fluids to each cassette.
- In some embodiments that cassettes will be connected in bricks of 10 or more cassettes, enabling large numbers of cassettes to be easily installed or removed from the system.
- The foregoing features of embodiments will be more readily understood by reference to the following detailed description, taken with reference to the accompanying drawings, in which:
-
FIG. 1 presents a cassette for culturing adherent cells comprising a first and second film; -
FIG. 2 presents a schematic diagram of an exemplary incubator that monitors and processes cell cultures disposed in cassettes; -
FIG. 3 depicts an exemplary incubator connected over a computer network to one or more remote servers; -
FIG. 4 presents the interior of an exemplary incubatory; -
FIG. 5 presents an exemplary incubator, -
FIG. 6 presents the interior of an exemplary table top model of an incubator; and -
FIG. 7 prevents an exemplary table top model of an incubator. - The invention relates to containers or cassettes for culturing adherent cells. The cassettes of the various embodiments offer numerous advantages over cell culture containers known in the art. For example, the cassettes and incubator permit automated detection of the cell culture environment, and automated passaging of cells, feeding of cells and sampling of cells. The cassettes also facilitate culture of large numbers of adherent cells in the presence of one or more compounds of interest. The cassettes allow for adherence of cells via methods that do not require chemical modification to detach the cells from the surface to which they adhere.
- The incubators have an increased capacity for housing cells in culture and are a designed to allow for automated monitoring and maintenance of the culture environment including but not limited to maintenance of pH, O2, CO2, and cell density. The incubators provide for automated sampling and imaging of cells in culture. The cassettes and incubator provide significant increases in the efficiency of methods of culturing adhesion cells.
- Definitions. As used in this description and the accompanying claims, the following terms shall have the meanings indicated, unless the context otherwise requires:
- As used herein, “cassette” means a sealed container for housing cells/and or tissues and culture media.
- As used herein, a “film” comprises any material suitable for containing cells, including but not limited to polymer films, for example, polyester, polystyrene, Surlyn, polyolefins, cyclic olefin copolymers (COC), or poly(methylmethacrylate) (PMMA), membranes, for example, PVDF, cellulose, or Nylon semi-permeable membranes. In one embodiment the film is metallized to minimize gas transfer
- In certain embodiments, cells adhere to a tie layer present on the film. In other embodiments, a film is coated with at least one compound including but not limited to a nutrient, a growth factor, a hormone, a cytokine, an antibiotic, an adhesion molecule, a mineral, an antibody, a nucleic acid, including but not limited to nucleic acid probes, primers and oligonucleotides, ligands, enzymes such as polymerases, proteases, isomerases, cell components or cell markers, small molecules and dyes. In certain embodiments, the compound is living or dead cells (e.g., for use as a viral target, or a baseline control for determining cell concentration or baseline or basal levels of target protein expression). The cells can be stained or treated to allow for detection. In certain embodiments, the “film” is coated with a test compound to be screened for a particular activity of interest. The film can be of any dimensions such that a surface for culturing the desired number of cells is provided. The thickness of the film can be such that diffusion of material across the surface of the film can occur. In one embodiment, the thickness of the film is from 1 μM to 1 mM, for example, 10 μM to 500 μM, or 20 μM to 100 μM.
- In one embodiment, the film comprises a tie layer(s) comprising a material that can be separated from or peeled off of the film.
- Cells adhere to the tie layer and therefore are removed from the film when the tie layer reverts to monomers or small polymers that dissolve upon exposure to a particular treatment, and wherein cells are subsequently released. For example, in one embodiment, a tie layer comprises a photo-depolymerizing material, or a material that, for example material that is thermoreversible, chemoreversible or enzymatically cleavable. Under the appropriate condition (light, heat, chemical environment) the tie layer reverts to monomers or small polymers that dissolve and release the cells. In one embodiment, film comprises a tie layer composed of a phase change material that allows the tie layer to be peeled off the film without reverting to monomers and small polymers that eventually dissolve, thereby providing a sheet of cells.
- As used herein, a “frame” refers to a structure that is used in association with the film comprising a cassette. A “frame” can be either solid or flexible. In certain embodiments, a “frame” comprises at least one sensor as defined herein. A “frame” is located either between a first and second film or on the exterior of a certain layer of film, for example, around the perimeter of the film. In certain embodiments, the frame comprises polystyrene, PMMA, polyolefins such as polyethylene, surlyn, engineering thermoplastics and any thermoplastic, thermoset or composite thereof.
- In certain embodiments, the film in combination with the frame is under tension.
- The cassette can include frangible sealed regions or compartments containing, for example, a liquid or solid where a sustained squeeze just prior to use will burst the seal and swirl the components together. The multiple component pouches with internal frangible seals afford the opportunity to conveniently and inexpensively deliver portions with time-of-use mixing. This allows for extending the shelf life of cassettes dramatically. All of the biologically active components are maintained in a fluid chamber that the user or machine can burst in. In one embodiment, the frangible film is Surlyn, which does not contribute particulars to the mixtures when the seal is broken.
- As used herein, a “sensor” detects or measures one or more properties of the cell and/or environment inside the cassette. A “sensor” can monitor, modify, record, indicate, or otherwise respond to one or more properties of the cell and/or the environment inside a cassette. According to the methods described herein, a sensor detects at least one of temperature, pH, O2, CO2 and cell density. In one embodiment, a sensor detects protein expression by detecting fluorescence, for example by a FISH assays. In other embodiments, the sensor is ligand-based or ligand-specific for detection of, for example, specific proteins or nucleic acids, apoptotic markers, nutrient levels, and/or the consumption rates of various media constituents.
- A sensor can be located on the film and/or the frame comprising the cassette.
- In certain embodiments, a “film” is coated with a compound useful for altering a property of a cell or tissue and/or supporting cell growth and viability and/or facilitating adhesion of the cells to the film. A “compound” includes but is not limited to a nutrient, a growth factor, a hormone, a cytokine, an antibiotic, an adhesion molecule, a mineral, an antibody, a nucleic acid, including a nucleic acid probe, an antisense molecule, a primer and an oligonucleotide, a ligand for a receptor of interest, enzymes, for example, including but not limited to polymerases, proteases, isomerases etc. . . . a small molecule and a dye. The compound can be a cell(s), either living or dead, for use as a viral target, for defining a baseline level of cellular activity and/or protein or mRNA expression; and to establish a baseline for cell concentration. The cells can be living or dead, and stained or otherwise treated to facilitate detection. The “film” can also be coated with a test compound to be screened for a particular activity of interest or a therapeutic agent.
- In certain embodiments, the cassette comprises a first and second layer of film, and further comprises one or more compartments. As used herein, a “compartment” refers to an enclosed section of the cassette, that can be used to house cells of interest such that they are separated from and are not in contact with cells that are present in the cassette but are located outside of the compartment. A “compartment” can include a sensor. The regions of the film comprising a compartment may be coated with at least one compound of interest. The compartment can comprise a particular cell media that is either different from or identical to media present in other compartments of the cassette. In certain embodiments, fluid is introduced to or removed from a “compartment” via a unique fluid line.
- The invention also provides for cassettes wherein replicate samples of cells are adhered to the film at discrete locations but are not confined to a “compartment” as defined herein.
- As used herein, “modulate” or “modulation” refers to increase or decrease.
- As used herein. “decrease” as it refers to the level of activity of a cell or the level of expression of a marker, for example a protein, mRNA or antibody of interest by a cell, means that activity or expression is 1, 2, 3, 4, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 100, 500, 1000 or 10,000-fold less before or after administration to or incubation in the presence of a cell with a compound of interest.
- As used herein, “decrease” as it refers to the level of activity of a cell or the level of expression of a marker, for example a protein, mRNA or antibody of interest by a cell, means that activity or expression is 1, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 99 or 100% before or after administration to or incubation in the presence of a cell with a compound of interest.
- As used herein “increased” as it refers to the level of activity of a cell or the level of expression of a marker, for example a protein, mRNA or antibody of interest by a cell, means that activity or expression is 1, 2, 3, 4, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 100, 500, 1000 or 10,000-fold or more before or after administration to or incubation in the presence of a cell with a compound of interest.
- As used herein “increased” as it refers to the level of activity of a cell or the level of expression of a marker, for example a protein. mRNA or antibody of interest by a cell is 1, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 99 or 100% greater before or after administration to or incubation in the presence of a cell with a compound of interest.
- As used herein, the term “disease” includes any one or more of the following autoimmune diseases or disorders: diabetes mellitus, arthritis (including rheumatoid arthritis, juvenile rheumatoid arthritis, osteoarthritis, psoriatic arthritis), multiple sclerosis, myasthenia gravis, systemic lupus erythematosis, autoimmune thyroiditis, dermatitis (including atopic dermatitis and eczematous dermatitis), psoriasis, Sjögren's Syndrome, including keratoconjunctivitis sicca secondary to Sjögren's Syndrome, alopecia areata, allergic responses due to arthropod bite reactions, Crohn's disease, aphthous ulcer, iritis, conjunctivitis, keratoconjunctivitis, ulcerative colitis, asthma, allergic asthma, cutaneous lupus erythematosus, scleroderma, vaginitis, proctitis, drug eruptions, leprosy reversal reactions, erythema nodosum leprosum, autoimmune uveitis, allergic encephalomyelitis, acute necrotizing hemorrhagic encephalopathy, idiopathic bilateral progressive sensorineural hearing loss, aplastic anemia, pure red cell anemia, idiopathic thrombocytopenia, polychondritis, Wegener's granulomatosis, chronic active hepatitis, Stevens-Johnson syndrome, idiopathic sprue, lichen planus, Graves ophthalmopathy, sarcoidosis, primary biliary cirrhosis, uveitis posterior, and interstitial lung fibrosis.
- In another embodiment, disease refers to any one of Wilson's disease, spinocerebellar ataxia, prion disease, Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis, amyloidosis, Alzheimer's disease, Alexander's disease, alcoholic liver disease, cystic fibrosis, Pick's Disease, spinal muscular dystrophy or Lewy body dementia.
- “Disease” also includes any one of rheumatoid spondylitis; post ischemic perfusion injury: inflammatory bowel disease; chronic inflammatory pulmonary disease, eczema, asthma, ischemia/reperfusion injury, acute respiratory distress syndrome, infectious arthritis, progressive chronic arthritis, deforming arthritis, traumatic arthritis, gouty arthritis, Reiter's syndrome, acute synovitis and spondylitis, glomerulonephritis, hemolytic anemia, aplastic anemia, neutropenia, host versus graft disease, allograft rejection, chronic thyroiditis. Graves' disease, primary binary cirrhosis, contact dermatitis, skin sunburns, chronic renal insufficiency, Guillain-Barre syndrome, uveitis, otitis media, periodontal disease, pulmonary interstitial fibrosis, bronchitis, rhinitis, sinusitis, pneumoconiosis, pulmonary insufficiency syndrome, pulmonary emphysema, pulmonary fibrosis, silicosis, or chronic inflammatory pulmonary disease.
- “Disease” also refers to any one of cancer, tumor growth, cancer of the colon, breast, bone, brain and others (e.g., osteosarcoma, neuroblastoma, colon adenocarcinoma), chronic myelogenous leukemia (CML), acute myeloid leukemia (AML), acute promyelocytic leukemia (APL), cardiac cancer (e.g., sarcoma, myxoma, rhabdomyoma, fibroma, lipoma and teratoma); lung cancer (e.g., bronchogenic carcinoma, alveolar carcinoma, bronchial adenoma, sarcoma, lymphoma, chondromatous hamartoma, mesothelioma): various gastrointestinal cancer (e.g., cancers of esophagus, stomach, pancreas, small bowel, and large bowel); genitourinary tract cancer (e.g., kidney, bladder and urethra, prostate, testis; liver cancer (e.g., hepatoma, cholangiocarcinoma, hepatoblastoma, angiosarcoma, hepatocellular adenoma, hemangioma); bone cancer (e.g., osteogenic sarcoma, fibrosarcoma, malignant fibrous histiocytoma, chondrosarcoma. Ewing's sarcoma, malignant lymphoma, multiple myeloma, malignant giant cell tumor chordoma, osteochronfroma, benign chondroma, chondroblastoma, chondromyxofibroma, osteoid osteoma and giant cell tumors); cancers of the nervous system (e.g., of the skull, meninges, brain, and spinal cord); gynecological cancers (e.g., uterus, cervix, ovaries, vulva, vagina); hematologic cancer (e.g., cancers relating to blood, Hodgkin's disease, non-Hodgkin's lymphoma); skin cancer (e.g., malignant melanoma, basal cell carcinoma, squamous cell carcinoma. Karposi's sarcoma, moles dysplastic nevi, lipoma, angioma, dermatofibroma, keloids, psoriasis); and cancers of the adrenal glands (e.g., neuroblastoma).
- As used herein, “diagnosing” or “identifying a patient or subject having” or “a patient or subject in need of treatment” refers to a process of determining if an individual is afflicted with a disease or ailment.
- A subject is said to be treated for a disease, if following administration of an agent, one or more symptoms of the disease are decreased or eliminated.
- “Treatment”, or “treating” as used herein, is defined as the application or administration of a compound identified as treating a disease of interest, to a subject or patient, or application or administration of a compound identified as treating a disease of interest to an isolated tissue or cell line from a subject or patient, who has a disease or disorder, with the purpose to cure, heal, alleviate, relieve, alter, remedy, ameliorate, improve or affect the disease or disorder, or symptoms of the disease or disorder. The term “treatment” or “treating” is also used herein in the context of administering agents prophylactically. The term “effective dose” or “effective amount” or “effective dosage” or “therapeutic dosage” is defined as an amount sufficient to achieve or at least partially achieve the desired effect. The terms “therapeutically effective dose” and “therapeutically effective amount” are defined as an amount sufficient to cure or at least partially arrest the disease and its complications in a patient already suffering from the disease.
- As used herein, “treating” a disease refers to preventing the onset of disease and/or reducing, delaying, or eliminating disease symptoms, such as an increase in the extracellular level of a group of cytokines. By “treating” is meant restoring the patient or subject to the basal state as defined herein, and/or to prevent a disease in a subject at risk thereof. Alternatively, “treating” means arresting or otherwise ameliorating symptoms of a disease.
- As used herein, “patient” or “subject” refers to a mammal that is diagnosed with a disease associated with an increase in the extracellular level of a group of cytokines.
- The term “patient” or “subject” includes human and other mammalian subjects that receive either prophylactic or therapeutic treatment.
- As used herein, “control subject” means a subject that has not been diagnosed with a disease and/or does not exhibit any detectable symptoms associated with the disease.
- As used herein, “mammal” refers to any mammal including but not limited to human, mouse, rat, sheep, monkey, goat, rabbit, hamster, horse, cow or pig.
- A “non-human mammal”, as used herein, refers to any mammal that is not a human.
- As used herein, “basal state” refers to the extracellular level of a group of cytokines of an individual who is not susceptible to a disease and who has no symptoms of a disease.
- As used herein “monitoring the treatment” means determining whether, following treatment of a subject, for example, a subject diagnosed with a disease associated with an increased extracellular level of a group of cytokines, the subject has been treated such that the symptoms of the disease are arrested or otherwise ameliorated and/or the disease and/or its attendant symptoms are alleviated or abated.
- As used herein, “pharmaceutically acceptable carrier” refers to a carrier for the administration of a therapeutic agent. Exemplary carriers include saline, buffered saline, dextrose, water, glycerol, ethanol, and combinations thereof.
- As used herein, the term “small molecule” refers to a non-peptidic, non-oligomeric organic compound either synthesized in the laboratory or found in nature. Small molecules, as used herein, can refer to compounds that are “natural product-like”. However, the term “small molecule” is not limited to “natural product-like” compounds. Rather, a small molecule is typically characterized in that it contains several carbon-carbon bonds, and has a molecular weight of less than 1500, although this characterization is not intended to be limiting for the purposes of the present invention. Examples of “small molecules” that occur in nature include, but are not limited to, taxol, dynemicin, and rapamycin. In certain other preferred embodiments, natural-product-like small molecules are utilized.
- The cassettes and incubator can be used to culture adherent cells under conditions that are precisely regulated. The cells can be maintained in culture by automated methods. The cassettes and incubator are useful for performing methods of treating a disease, methods of diagnosing a disease, methods of monitoring disease treatment and methods of designing a treatment protocol. In addition, the cassettes and incubator facilitate screening for a compound, for example, a therapeutic.
- Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, suitable methods and materials are described below. All publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety. In case of conflict, the present specification, including definitions, will control. In addition, the materials, methods, and examples are illustrative only and not intended to be limiting.
-
FIG. 1 presents one embodiment of a cassette comprising a first and second layer of film and a frame. The cassette can be prepared by a method including preparing a frame that is, for example, injected molded, comprises a laminate-layered core or cast core or center core, is 3D printed, machined or lasercut. In one embodiment, the frame is surrounded on the top and bottom by film, membrane, multilayer coated film or a combination thereof. In another embodiment, the frame is surround on the top and bottom with a thick film that is die-stamped, laser cut, etc. . . . - The film is coated at least by any of the following methods: roll to roll coating, printing of certain regions of the film, for example PRP islets to which discrete populations of cells will adhere, reagents used for detection, spots, lanes or other structures, hydrophobic and hydrophilic regions to channel liquid flow within the cassette.
- In other embodiments, the file is coated with a metallic substance or other vapor or gas control coatings, for example by chemical vapor deposit (CVD) or sputtering, gravure coating, skiving and any other method of applying a coating reagent, ink or dye to a film or surface known in the art.
- In one embodiment, cells are introduced into a cassette using a sterilizable and/or disposable, microfluidic manifold located in the rear of the chamber that controls the flow of median to each of the cassettes. For automated introduction, a micro plate is inserted into the machine, and the cells are individually aspirated from the plate via a disposable or reusable sterilizable capillary device that interfaces with the manifold. The capillary is robotically moved or and through gates and microfluidics the flow can be directed. For example, a robot that traverses the entire back of the system, has a capillary injection port that butts against the array or the cassettes themselves and injects the cells. Movement of media in an out the cassette can occur via this process.
- For manual injection the user can lift a corner of the film and inject via pipet (such a liftable corner is depicted in
FIG. 1 ) then reseal (for example, via an adhesive); or the user can inject through a re-sealable port that is built into the film or frame or both of the cassette. -
FIG. 2 represents a schematic diagram of anexemplary incubator 200 that monitors and processes cell cultures disposed in cassettes. Theincubator 200 includes aprogrammable computing device 205 that controls the incubator's operations. Thecomputing device 205 includes a user interface, a processor, and a memory. The memory stores instructions for operating theincubator 200. Thecomputing device 205 receives parameters from a user for processing cell cultures. Thecomputing device 205 executes instructions according to the parameters to control components of theincubator 200. In various embodiments, thecomputing device 205 is configured to communicate overcomputer networks 305 withremote servers 310. Theremote servers 310 may store data from thecomputing device 205 or communicate with users according to instructions from thecomputing device 205, as described in more detail below. - The incubator includes a
chassis 210 disposed in ahousing 211. Thechassis 210 includes shelves that receive and support cassettes containing cell cultures. The shelves have a plurality ofports computing device 205. The shelf's port 215 interfaces with a connector on the cassette that is coupled to the cassette's circuitry, which receives data from sensors disposed within the cassette. The port 215 includes one or more connections for supplying power to the cassette's circuitry and/or receiving data collected by the circuitry's sensors. Data received through this port 215 is transmitted to thecomputing device 205. - The
incubator 200 includes afluidic manifold 220. Afluidic manifold 220 has achamber 225 that houses a media of interest useful for cell culture. In some embodiments, thechamber 225 receives cell cultures extracted from cassettes. Further, although the figure depicts asingle chamber 225, various embodiments of theincubator 200 may include multiple chambers for housing different media, or multiple chambers for receiving cell cultures from different cassettes. In this embodiment, aconduit 230 extends from thechamber 225 to shelves in thechassis 210. Theconduit 230 includesnodes - In certain embodiments, a
pump 240 coupled to thefluidic manifold 220 can propel media in thechamber 225 through a conduit into one or more of the cassettes. Tn further embodiments, avacuum 245 coupled to thefluidic manifold 220 extracts the contents of one or more cassettes into a chamber. In some embodiments, thefluidic manifold 220 extracts the contents of a cassette into the same chamber used for propelling media into cassettes in the incubator. In other embodiments, thefluidic manifold 220 includes separate chambers for housing media to propel into cassettes and receiving contents extracted from cassettes. - In some embodiments, the
incubator 200 has one or more image capturing devices and robotic arms for positioning the devices within thehousing 211. Thecomputing device 205 operates the robotic arms to position an image capturing device over a cell culture in a cassette. Thecomputing device 205 operates the image capturing device to obtain an image of a cell culture. -
FIG. 3 depicts theincubator 200 connected over acomputer network 305 to one or moreremote servers 310. Thecomputing device 205 connects to the remote server(s) 310 via a local area connection (LAN), wide area connection (WAN), the Internet, or any computer network. In some embodiments, thecomputing device 205 transmits data regarding a cell culture, for example, pH and/or cell density, and environmental conditions within cassettes to aremote server 310 for storage. The server 315 stores the data. In some embodiments, thecomputing device 205 transmits messages to a remote server 315 for communicating with a user of the incubator. In turn, the remote server 315 sends the messages to a user. - In operation, a user introduces cells in combination with a media of interest into the cassettes and inserts the cassettes into the shelves of the
incubator 200. When the user connects the connector on the cassette with the shelf's port 215, thecomputing device 205 detects the presence of the cassette. In some embodiments, thecomputing device 205 powers the cassette's circuitry by supplying a power signal through the port 215. Thecomputing device 205 initializes a file for storing data on the environmental conditions of the cassette. Thecomputing device 205 receives data collected by sensors in the cassette, such as temperature, cell density, and pH. In some embodiments, thecomputing device 205 stores the data in the file associated with the cassette. In other embodiments, thecomputing device 205 discards the data until thedevice 205 receives processing instructions from the user. - A user programs the
computing device 205 to process cell cultures disposed in cassettes in theincubator 200. In some embodiments, the user programs thecomputing device 205 from a user interface disposed on theincubator 200. The user interface displays a menu of parameters for the user to select. For example, the user can select the temperature and humidity at which the incubator will maintain the environment in the cassettes. The user can select the media to introduce into the cassettes during processing. In certain embodiments, the user interface displays the % concentration of a gas, for example, O2 or CO2, the temperature inside the incubator, the composition and/or volume and/or pH of the media, or the amount of other substances present in the incubator's chambers, and the user can supply the incubator with additional resources, if necessary. - The user can create a schedule of actions with respect to the cell cultures. Some of the actions may be directed to monitoring the cell cultures. For example, the user may configure the incubator to begin beeping every 6 hours, thereby alerting the user to check conditions of the cell cultures. In another example, the user may configure the incubator to send the user a short message service (SMS) communication every 6 hours, for the same purpose. The user may input a mobile telephone number at which the user is capable of receiving SMS communication. Every six hours, the
computing device 205 sends an instruction to theremote server 310 to send a reminder message to the user s mobile telephone number. In response, theremote server 310 creates the corresponding SMS communication and sends the same to the telephone number. - In a similar example, the user may configure the incubator to capture an image of the cells present in a particular cassette and e-mail the image to the user every 6 hours. The user may input an e-mail address. Every six hours, the
computing device 205 operates a robotic arm to position an image capturing device over the cassette of interest, capture an image, and send the image and e-mail address to aremote server 310. Theremote server 310 creates an e-mail message with the image and sends the same to the user. - Some of the actions may be directed to maintaining environmental conditions within the cassette. For example, the user can select a desired pH level for a cell culture, an acceptable level of deviation from the desired pH level, and media for adjusting the pH level in a cassette. Thus, if the pH level in a cassette becomes too basic, the
fluidic manifold 220 flows additional media of the user's choosing into the cassette. Likewise, thefluidic manifold 220 flows a different media selected by the user if the pH level becomes too acidic. In another example, the user can select a desired level of feed for the cell culture. For example, the user can select a desired concentration, or a minimum concentration, of feed present in the cell culture's environment. When the concentration of feed drops below the desired or minimum level, thefluidic manifold 220 flows additional feed into the cassette. - Some of the actions may be directed to processing a cell culture within a cassette. An action may depend upon elapses of time, conditions regarding the cell culture itself or the environment in the cassette (e.g., pH, temperature, cell density), or user initiative. In various examples, the user may schedule the incubator to flow a media into ten cassettes comprising cell cultures, heat the cassettes until they reach a desired temperature, and maintain the desired temperature for a particular length of time. The user may schedule the incubator to monitor the pH levels of the cassettes and flow a selected media through the cassettes once the pH levels reach a user selected threshold. The user may schedule the incubator to monitor the cell density of cultures and passage the same once the cell density reaches a desired threshold. The
fluidic manifold 220 flows the passaging media into the cassettes, and the vacuum extracts the cells into a chamber for retrieval by the user. - In some embodiments, the user may schedule the incubator to send alerts whenever a programmed condition is met. For example, a user may schedule the incubator to flow a media into cassettes when the pH level in the cassette drops below a threshold level. The
computing device 205 continues to receive data about the pH levels from the cassettes, and when the pH levels drop to the threshold, thecomputing device 205 sends the user an alert (e.g., an e-mail message, an SMS message). Thus, the user may be informed of an experiment's progress. - In some embodiments, the user may schedule the incubator to monitor the cell density and send the user an image of a culture when the detected cell density reaches a desired threshold. The incubator may require user approval to passage the cells, or to enact another process. If the incubator does not receive such approval, the incubator may maintain the environmental conditions of the cassette and continue sending images to the user, according to a predetermined schedule. Once the user approves an image of the cell cultures, based on its density, the incubator may passage the cell culture, or perform some other desired process.
- Processing actions may include any manipulation of the cell cultures, according to any conditions. The user can schedule a series of processes, based on any of the parameters described herein. For example, the user can instruct the incubator to flow medium A into the cassettes and allow the cells to incubate for a particular period of time. After the four hours have elapsed, the incubator may flow medium B into the cassettes until the pH level for each cassette has dropped to a user selected level. Then, the incubator may raise the temperature of the incubator to a desired temperature. Once the temperature reaches the desired temperature, the fluidic manifold may flow medium C into the cassettes to feed the cell cultures. The incubator may monitor the cell density of each cassette. When the cell density of a cassette reaches a user selected threshold, the fluidic manifold may flow a passaging reagent into the cassettes, the cells are incubated for a predetermined period of time in the presence of the reagent such that the cells dissociate from the film comprising the cassette, and vacuum the cassettes' contents into a chamber from which the cells can be isolated. Processes according to similar parameters, or sequences thereof, may also be deployed.
- In some embodiments, the user can program the
computing device 205 remotely. The user can log into aremote computing device 205 that executes an application that instructs the sequence of events required to perform a particular experimental analysis. The application may include a similar user interface to the one depicted on the incubator. The user may create a schedule of actions for the incubator, as described herein. Thecomputing device 205 executing the application transmits the user controls over acomputer network 305 to the incubator, and the incubator'scomputing device 205 stores the controls in its memory. - Once the user inputs the schedule of actions, the
computing device 205 receives data about environmental conditions in the cassettes through the ports 215 and store the data in files corresponding to the cassettes. In some embodiments, thecomputing device 205 stores the data in its local memory. The user can access the data through the user interface. From the menu, the user can select the file corresponding to a cassette of interest, and thecomputing device 205 displays the cassette's data. The user can scroll through the data to view data at different intervals of time. - In other embodiments, the
computing device 205 transmits the data over acomputer network 305 to a remote server for storage. Users can access the data on theremote servers 310. In some embodiments, using an application executing on a computing device, a user can log into an account on theremote server 310. The user may view the data collected by theincubator 200 and transmitted to theremote server 310. In this manner, the user may monitor the progress of the cell cultures and the environmental conditions through which the cell cultures have progressed. - Various embodiments may be implemented at least in part in any conventional computer programming language. For example, some embodiments may be implemented in a procedural programming language (e.g., “C”), or in an object oriented programming language (e.g., “C++”). Other embodiments may be implemented as preprogrammed hardware elements (e.g., application specific integrated circuits, FPGAs. and digital signal processors), or other related components.
- In an alternative embodiment, the disclosed apparatus and methods may be implemented as a computer program product for use with a computer system. Such implementation may include a series of computer instructions fixed either on a tangible medium, such as a computer readable medium (e.g., a diskette, CD-ROM. ROM, or fixed disk). The series of computer instructions can embody all or part of the functionality previously described herein with respect to the system.
- Those skilled in the art should appreciate that such computer instructions can be written in a number of programming languages for use with many computer architectures or operating systems. Furthermore, such instructions may be stored in any memory device, such as semiconductor, magnetic, optical or other memory devices, and may be transmitted using any communications technology, such as optical, infrared, microwave, or other transmission technologies.
- Among other ways, such a computer program product may be distributed as a tangible removable medium with accompanying printed or electronic documentation (e.g., shrink wrapped software), preloaded with a computer system (e.g., on system ROM or fixed disk), or distributed from a server or electronic bulletin board over the network (e.g., the Internet or World Wide Web). Of course, some embodiments may be implemented as a combination of both software (e.g., a computer program product) and hardware. Still other embodiments are implemented as entirely hardware, or entirely software.
- A cell of interest is introduced into a cassette of the invention via a sterilizable and/or disposable microfluidic manifold located in the rear of the chamber that controls the follow of medium to a cassette of the invention. A cell is introduced either manually or via automated methods. A cell is introduced in combination with a media of interest into a cassette and the cassette is inserted into an incubator according to the invention. In certain embodiments, the cell is cultured in the presence of at least one agent known to have a desired effect on the cell, for example, increase or decrease an activity of the cell, or increase or decrease expression of a marker of the cell. The cell can also be cultured in the presence of an agent identified as treating a disease. Cells are cultured at a density know to be appropriate for optimal growth of that particular cell type.
- Cells are removed from a subject in need of treatment. Cells that are removed can be present in a biological sample including but not limited to, blood, saliva, urine and tissue. The cells are transferred into the cassette of the invention and cultured in the presence of at least one agent identified as treating the disease. Following the culture period, the cells are reintroduced into the subject, for example, by injection, thereby treating the disease.
- Various embodiments of the present invention may be characterized by the potential claims listed in the paragraphs following this paragraph (and before the actual claims provided at the end of this application). These potential claims form a part of the written description of this application. Accordingly, subject matter of the following potential claims may be presented as actual claims in later proceedings involving this application or any application claiming priority based on this application. Inclusion of such potential claims should not be construed to mean that the actual claims do not cover the subject matter of the potential claims. Thus, a decision to not present these potential claims in later proceedings should not be construed as a donation of the subject matter to the public.
- The embodiments described above are intended to be merely exemplary; numerous variations and modifications will be apparent to those skilled in the art. All such variations and modifications are intended to be within the scope of the present invention as defined in any appended claims.
Claims (7)
1. A method of operating an incubator, the method comprising:
storing, by a computing device, parameters for a schedule of processes to apply to cell cultures in cassettes;
placing the cell culture cassettes into the incubator for cell culturing;
imaging cell cultures in the cell culture cassettes;
monitoring, by the computing device, data about environmental conditions and cell density during cell culturing;
sending a user an image of the culture when the monitored density reaches a desired threshold;
passaging the cells or performing another process in response to a user input regarding the image of the culture.
2. The method of operating the incubator of claim 1 , further comprising:
receiving data about environmental conditions at a remote server from the.
3. The method according to claim 1 , wherein the image is transmitted to a remote server and the remote server emails the image to the user.
4. The method according to claim 3 , wherein the imager transmits the image to the remote server according to a time schedule.
5. The method according to claim 1 , wherein the step of imaging comprises positioning an imaging device over the cell culture using a robotic arm.
6. The method according to claim 1 , further comprising sending an alert to the user to check environmental conditions.
7. The method according to claim 6 , wherein the alert is an SMS communication.
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US17/505,209 US20220106549A1 (en) | 2013-03-15 | 2021-10-19 | Cell culture cassettes and incubator |
Applications Claiming Priority (6)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US201361788745P | 2013-03-15 | 2013-03-15 | |
US201361802976P | 2013-03-18 | 2013-03-18 | |
US14/211,711 US9597355B2 (en) | 2013-03-15 | 2014-03-14 | Cell culture cassettes and incubator |
US15/427,887 US10519411B2 (en) | 2013-03-15 | 2017-02-08 | Cell culture cassettes and incubator |
US16/694,285 US20200087607A1 (en) | 2013-03-15 | 2019-11-25 | Cell culture cassettes and incubator |
US17/505,209 US20220106549A1 (en) | 2013-03-15 | 2021-10-19 | Cell culture cassettes and incubator |
Related Parent Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US16/694,285 Continuation US20200087607A1 (en) | 2013-03-15 | 2019-11-25 | Cell culture cassettes and incubator |
Publications (1)
Publication Number | Publication Date |
---|---|
US20220106549A1 true US20220106549A1 (en) | 2022-04-07 |
Family
ID=51527931
Family Applications (4)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US14/211,711 Active US9597355B2 (en) | 2013-03-15 | 2014-03-14 | Cell culture cassettes and incubator |
US15/427,887 Active 2034-05-27 US10519411B2 (en) | 2013-03-15 | 2017-02-08 | Cell culture cassettes and incubator |
US16/694,285 Abandoned US20200087607A1 (en) | 2013-03-15 | 2019-11-25 | Cell culture cassettes and incubator |
US17/505,209 Pending US20220106549A1 (en) | 2013-03-15 | 2021-10-19 | Cell culture cassettes and incubator |
Family Applications Before (3)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US14/211,711 Active US9597355B2 (en) | 2013-03-15 | 2014-03-14 | Cell culture cassettes and incubator |
US15/427,887 Active 2034-05-27 US10519411B2 (en) | 2013-03-15 | 2017-02-08 | Cell culture cassettes and incubator |
US16/694,285 Abandoned US20200087607A1 (en) | 2013-03-15 | 2019-11-25 | Cell culture cassettes and incubator |
Country Status (2)
Country | Link |
---|---|
US (4) | US9597355B2 (en) |
WO (1) | WO2014143737A1 (en) |
Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US11643667B2 (en) | 2017-08-28 | 2023-05-09 | Cellino Biotech, Inc. | Microfluidic laser-activated intracellular delivery systems and methods |
US11680247B2 (en) | 2021-03-07 | 2023-06-20 | Cellino Biotech, Inc. | Platforms and systems for automated cell culture |
US11931737B2 (en) | 2021-09-02 | 2024-03-19 | Cellino Biotech, Inc. | Platforms and systems for automated cell culture |
Families Citing this family (15)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP6167309B2 (en) * | 2015-08-03 | 2017-07-26 | パナソニックIpマネジメント株式会社 | Method for determining whether a test sample contains phytopathogenic oomycete |
US11308121B2 (en) | 2015-11-04 | 2022-04-19 | Thrive Bioscience, Inc. | Method and system for cell operations using a blockchain data network |
US11230690B2 (en) | 2015-11-04 | 2022-01-25 | Thrive Bioscience, Inc. | Method and system for cell operations using the internet of things |
WO2017079692A1 (en) | 2015-11-04 | 2017-05-11 | Thrive Bioscience, Inc. | Networked incubator operation |
JP6995369B2 (en) * | 2015-11-18 | 2022-01-14 | スライブ バイオサイエンス, インコーポレイテッド | Instrument resource scheduling |
EP3504315A4 (en) | 2016-08-27 | 2020-04-15 | 3D Biotek, LLC | Bioreactor |
EP3404090A1 (en) * | 2017-05-15 | 2018-11-21 | Eppendorf AG | Incubator, system and method for monitored cell growth |
CN108693004B (en) * | 2018-05-24 | 2020-08-04 | 中国农业科学院农业质量标准与检测技术研究所 | Method for detecting whether rapeseed meal is doped with antibiotic filter residues or not |
CN108841036B (en) * | 2018-07-13 | 2019-04-19 | 四川大学 | A kind of CO2Responsiveness ordered porous membrane material and preparation method thereof |
CN113316724A (en) | 2018-11-16 | 2021-08-27 | 爱新诺有限公司 | Processing module for automated biological systems |
JP7333818B2 (en) | 2018-11-16 | 2023-08-25 | アイキシンノ・リミテッド | Module for automated biological laboratory systems with interfaces for transferring microplates or labware |
JP7303300B2 (en) | 2018-11-16 | 2023-07-04 | アイキシンノ・リミテッド | Labware handling system for labware and cell culture processes |
US20220010254A1 (en) | 2018-11-16 | 2022-01-13 | Aixinno Limited | A system for processing biology material, comprising a plurality of uniform design storage modules |
US20230108890A1 (en) * | 2020-02-10 | 2023-04-06 | Integriculture Inc. | Cell culture management system |
WO2021183687A2 (en) | 2020-03-10 | 2021-09-16 | Cellares Corporation | Systems, devices, and methods for cell processing |
Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20020146817A1 (en) * | 2000-10-02 | 2002-10-10 | Cannon Thomas F. | Automated bioculture and bioculture experiments system |
Family Cites Families (13)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US1881118A (en) * | 1930-03-01 | 1932-10-04 | Clair R Bratton | Incubator |
US20020110905A1 (en) * | 2001-02-15 | 2002-08-15 | Emilio Barbera-Guillem | Perfusion system for cultured cells |
WO2005059088A1 (en) * | 2003-12-19 | 2005-06-30 | University Of Waterloo | Cultured cell and method and apparatus for cell culture |
EP1869476A2 (en) * | 2005-03-22 | 2007-12-26 | Irm Llc | Compound profiling devices, systems, and related methods |
EP1944358B1 (en) * | 2005-11-01 | 2017-08-02 | Medinet., Co. Ltd. | Cell culture apparatus, cell culture method, cell culture program and cell culture system |
EP2076774A1 (en) * | 2007-02-28 | 2009-07-08 | Corning Incorporated | Surfaces and methods for biosensor cellular assays |
EP2130905A1 (en) | 2008-06-04 | 2009-12-09 | Pharmacell B.V. | Method for culturing eukaryotic cells |
WO2010042072A1 (en) * | 2008-10-08 | 2010-04-15 | Agency For Science, Technology And Research | Apparatus for culturing anchorage dependent cells |
EP2192410B1 (en) | 2008-11-26 | 2012-07-11 | Corning Incorporated | Nanoparticulate affininty capture for label independent detections system |
WO2010069320A2 (en) * | 2008-12-19 | 2010-06-24 | Stobbe Tech A/S | Biopharmaceutical plant in a column |
US7956328B2 (en) * | 2009-07-29 | 2011-06-07 | Battelle Memorial Institute | System, device, and methods for real-time screening of live cells, biomarkers, and chemical signatures |
US9550970B2 (en) * | 2010-02-17 | 2017-01-24 | Inq Biosciences Corporation | Culture systems, apparatus, and related methods and articles |
TW201137121A (en) * | 2010-04-26 | 2011-11-01 | Univ Nat Changhua Education | Cell culture real-time observation system |
-
2014
- 2014-03-14 US US14/211,711 patent/US9597355B2/en active Active
- 2014-03-14 WO PCT/US2014/027821 patent/WO2014143737A1/en active Application Filing
-
2017
- 2017-02-08 US US15/427,887 patent/US10519411B2/en active Active
-
2019
- 2019-11-25 US US16/694,285 patent/US20200087607A1/en not_active Abandoned
-
2021
- 2021-10-19 US US17/505,209 patent/US20220106549A1/en active Pending
Patent Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20020146817A1 (en) * | 2000-10-02 | 2002-10-10 | Cannon Thomas F. | Automated bioculture and bioculture experiments system |
Non-Patent Citations (1)
Title |
---|
Ker, D.F.E. et al. 2011. An engineered approach to stem cell culture: automating the decision process for real-time adaptive subculture of stem cells. PLoS ONE 6(11): 1-12; specif. pp. 1, 2, 7, 10 * |
Cited By (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US11643667B2 (en) | 2017-08-28 | 2023-05-09 | Cellino Biotech, Inc. | Microfluidic laser-activated intracellular delivery systems and methods |
US11680247B2 (en) | 2021-03-07 | 2023-06-20 | Cellino Biotech, Inc. | Platforms and systems for automated cell culture |
US11708563B2 (en) | 2021-03-07 | 2023-07-25 | Cellino Biotech, Inc. | Platforms and systems for automated cell culture |
US11866735B2 (en) | 2021-03-07 | 2024-01-09 | Cellino Biotech, Inc. | Platforms and systems for automated cell culture |
US11913029B2 (en) | 2021-03-07 | 2024-02-27 | Cellino Biotech, Inc. | Platforms and systems for automated cell culture |
US11931737B2 (en) | 2021-09-02 | 2024-03-19 | Cellino Biotech, Inc. | Platforms and systems for automated cell culture |
Also Published As
Publication number | Publication date |
---|---|
WO2014143737A1 (en) | 2014-09-18 |
US20170145366A1 (en) | 2017-05-25 |
US20200087607A1 (en) | 2020-03-19 |
US20140271571A1 (en) | 2014-09-18 |
US9597355B2 (en) | 2017-03-21 |
US10519411B2 (en) | 2019-12-31 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
US20220106549A1 (en) | Cell culture cassettes and incubator | |
Sung et al. | Recent advances in body-on-a-chip systems | |
Banaeiyan et al. | Design and fabrication of a scalable liver-lobule-on-a-chip microphysiological platform | |
Hung et al. | Continuous perfusion microfluidic cell culture array for high‐throughput cell‐based assays | |
Agastin et al. | Continuously perfused microbubble array for 3D tumor spheroid model | |
Zhang et al. | A robust high-throughput sandwich cell-based drug screening platform | |
Underhill et al. | Advances in engineered human liver platforms for drug metabolism studies | |
He et al. | Hierarchical spiky microstraws‐integrated microfluidic device for efficient capture and in situ manipulation of cancer cells | |
US10988723B1 (en) | Modular assemblies and systems for cell cultures and methods thereof | |
US8828332B2 (en) | Microfluidic capsule | |
EP2831221A1 (en) | Cell culture apparatus and culture methods using same | |
Arakawa et al. | High-throughput single-cell manipulation system for a large number of target cells | |
Hosic et al. | Rapid prototyping of multilayer microphysiological systems | |
Orbach et al. | In vitro intestinal and liver models for toxicity testing | |
US9834747B2 (en) | Methods and apparatus for transplantation of nucleic acid molecules | |
Liu et al. | Large-scale antitumor screening based on heterotypic 3D tumors using an integrated microfluidic platform | |
Tang et al. | On-Chip cell–cell interaction monitoring at single-cell level by efficient immobilization of multiple cells in adjustable quantities | |
Lin et al. | Orthogonal screening of anticancer drugs using an open-access microfluidic tissue array system | |
Mughal et al. | Organs‐on‐Chips: Trends and Challenges in Advanced Systems Integration | |
Feng et al. | Antibody-free isolation and regulation of adherent cancer cells via hybrid branched microtube-sandwiched hydrodynamic system | |
CN101376908B (en) | Method for studying medicament metabolism based on molecule and cell level | |
Zhou et al. | Degradable porous nanoflower substrate-embedded microfluidic device for capture, release and in situ manipulation of cancer cells | |
Parihar et al. | Tumor-on-a-chip: microfluidic models of hypoxic tumor microenvironment | |
CN116445284A (en) | Tumor cell-vascular endothelial cell in-vitro co-culture device and preparation method and application thereof | |
Macdonald et al. | Creating tissue on chip constructs in microtitre plates for drug discovery |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
STPP | Information on status: patent application and granting procedure in general |
Free format text: DOCKETED NEW CASE - READY FOR EXAMINATION |
|
STPP | Information on status: patent application and granting procedure in general |
Free format text: NON FINAL ACTION MAILED |
|
STPP | Information on status: patent application and granting procedure in general |
Free format text: RESPONSE TO NON-FINAL OFFICE ACTION ENTERED AND FORWARDED TO EXAMINER |
|
STPP | Information on status: patent application and granting procedure in general |
Free format text: NON FINAL ACTION MAILED |
|
STPP | Information on status: patent application and granting procedure in general |
Free format text: RESPONSE TO NON-FINAL OFFICE ACTION ENTERED AND FORWARDED TO EXAMINER |
|
STPP | Information on status: patent application and granting procedure in general |
Free format text: FINAL REJECTION MAILED |