CN117122668B - 一种具有改善呼吸系统感染作用的全阵列细胞因子及其制备方法 - Google Patents
一种具有改善呼吸系统感染作用的全阵列细胞因子及其制备方法 Download PDFInfo
- Publication number
- CN117122668B CN117122668B CN202310630711.6A CN202310630711A CN117122668B CN 117122668 B CN117122668 B CN 117122668B CN 202310630711 A CN202310630711 A CN 202310630711A CN 117122668 B CN117122668 B CN 117122668B
- Authority
- CN
- China
- Prior art keywords
- cells
- cell
- cytokine
- primary
- medium
- 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.)
- Active
Links
- 102000004127 Cytokines Human genes 0.000 title claims abstract description 39
- 108090000695 Cytokines Proteins 0.000 title claims abstract description 39
- 230000000694 effects Effects 0.000 title claims abstract description 12
- 238000002360 preparation method Methods 0.000 title abstract description 7
- 210000002345 respiratory system Anatomy 0.000 title description 7
- 208000015181 infectious disease Diseases 0.000 title description 5
- 102100021943 C-C motif chemokine 2 Human genes 0.000 claims abstract description 6
- 206010057190 Respiratory tract infections Diseases 0.000 claims abstract description 5
- 102000005789 Vascular Endothelial Growth Factors Human genes 0.000 claims abstract description 4
- 108010019530 Vascular Endothelial Growth Factors Proteins 0.000 claims abstract description 4
- MZOFCQQQCNRIBI-VMXHOPILSA-N (3s)-4-[[(2s)-1-[[(2s)-1-[[(1s)-1-carboxy-2-hydroxyethyl]amino]-4-methyl-1-oxopentan-2-yl]amino]-5-(diaminomethylideneamino)-1-oxopentan-2-yl]amino]-3-[[2-[[(2s)-2,6-diaminohexanoyl]amino]acetyl]amino]-4-oxobutanoic acid Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CC(O)=O)NC(=O)CNC(=O)[C@@H](N)CCCCN MZOFCQQQCNRIBI-VMXHOPILSA-N 0.000 claims abstract description 3
- 108010081589 Becaplermin Proteins 0.000 claims abstract description 3
- 101710155857 C-C motif chemokine 2 Proteins 0.000 claims abstract description 3
- 102100032367 C-C motif chemokine 5 Human genes 0.000 claims abstract description 3
- 108700012434 CCL3 Proteins 0.000 claims abstract description 3
- 102000000013 Chemokine CCL3 Human genes 0.000 claims abstract description 3
- 102000001326 Chemokine CCL4 Human genes 0.000 claims abstract description 3
- 108010055165 Chemokine CCL4 Proteins 0.000 claims abstract description 3
- 108010055166 Chemokine CCL5 Proteins 0.000 claims abstract description 3
- 102000004269 Granulocyte Colony-Stimulating Factor Human genes 0.000 claims abstract description 3
- 108010017080 Granulocyte Colony-Stimulating Factor Proteins 0.000 claims abstract description 3
- 108010017213 Granulocyte-Macrophage Colony-Stimulating Factor Proteins 0.000 claims abstract description 3
- 102100039620 Granulocyte-macrophage colony-stimulating factor Human genes 0.000 claims abstract description 3
- 101000897480 Homo sapiens C-C motif chemokine 2 Proteins 0.000 claims abstract description 3
- 102100037850 Interferon gamma Human genes 0.000 claims abstract description 3
- 108010074328 Interferon-gamma Proteins 0.000 claims abstract description 3
- 108050003558 Interleukin-17 Proteins 0.000 claims abstract description 3
- 102000013691 Interleukin-17 Human genes 0.000 claims abstract description 3
- 108010002350 Interleukin-2 Proteins 0.000 claims abstract description 3
- 108090000978 Interleukin-4 Proteins 0.000 claims abstract description 3
- 108010002616 Interleukin-5 Proteins 0.000 claims abstract description 3
- 108090001005 Interleukin-6 Proteins 0.000 claims abstract description 3
- 108010002586 Interleukin-7 Proteins 0.000 claims abstract description 3
- 108090001007 Interleukin-8 Proteins 0.000 claims abstract description 3
- 108010002335 Interleukin-9 Proteins 0.000 claims abstract description 3
- 108060008682 Tumor Necrosis Factor Proteins 0.000 claims abstract description 3
- 102000000852 Tumor Necrosis Factor-alpha Human genes 0.000 claims abstract description 3
- 102000009634 interleukin-1 receptor antagonist activity proteins Human genes 0.000 claims abstract description 3
- 108040001669 interleukin-1 receptor antagonist activity proteins Proteins 0.000 claims abstract description 3
- 210000004027 cell Anatomy 0.000 claims description 100
- NOESYZHRGYRDHS-UHFFFAOYSA-N insulin Chemical compound N1C(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(NC(=O)CN)C(C)CC)CSSCC(C(NC(CO)C(=O)NC(CC(C)C)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CCC(N)=O)C(=O)NC(CC(C)C)C(=O)NC(CCC(O)=O)C(=O)NC(CC(N)=O)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CSSCC(NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2C=CC(O)=CC=2)NC(=O)C(CC(C)C)NC(=O)C(C)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2NC=NC=2)NC(=O)C(CO)NC(=O)CNC2=O)C(=O)NCC(=O)NC(CCC(O)=O)C(=O)NC(CCCNC(N)=N)C(=O)NCC(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC(O)=CC=3)C(=O)NC(C(C)O)C(=O)N3C(CCC3)C(=O)NC(CCCCN)C(=O)NC(C)C(O)=O)C(=O)NC(CC(N)=O)C(O)=O)=O)NC(=O)C(C(C)CC)NC(=O)C(CO)NC(=O)C(C(C)O)NC(=O)C1CSSCC2NC(=O)C(CC(C)C)NC(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CC(N)=O)NC(=O)C(NC(=O)C(N)CC=1C=CC=CC=1)C(C)C)CC1=CN=CN1 NOESYZHRGYRDHS-UHFFFAOYSA-N 0.000 claims description 30
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 26
- 210000003954 umbilical cord Anatomy 0.000 claims description 19
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 claims description 16
- 102000004877 Insulin Human genes 0.000 claims description 16
- 108090001061 Insulin Proteins 0.000 claims description 16
- DAEPDZWVDSPTHF-UHFFFAOYSA-M sodium pyruvate Chemical compound [Na+].CC(=O)C([O-])=O DAEPDZWVDSPTHF-UHFFFAOYSA-M 0.000 claims description 16
- 239000012879 subculture medium Substances 0.000 claims description 16
- 102100037362 Fibronectin Human genes 0.000 claims description 15
- 108010067306 Fibronectins Proteins 0.000 claims description 15
- 229940125396 insulin Drugs 0.000 claims description 15
- HVYWMOMLDIMFJA-DPAQBDIFSA-N cholesterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 HVYWMOMLDIMFJA-DPAQBDIFSA-N 0.000 claims description 14
- 238000004108 freeze drying Methods 0.000 claims description 13
- 239000001963 growth medium Substances 0.000 claims description 13
- 239000007788 liquid Substances 0.000 claims description 13
- 229910052757 nitrogen Inorganic materials 0.000 claims description 13
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 13
- 238000012258 culturing Methods 0.000 claims description 10
- 239000002609 medium Substances 0.000 claims description 10
- 238000000034 method Methods 0.000 claims description 10
- 239000006228 supernatant Substances 0.000 claims description 10
- 238000007710 freezing Methods 0.000 claims description 9
- 230000008014 freezing Effects 0.000 claims description 9
- 210000002901 mesenchymal stem cell Anatomy 0.000 claims description 9
- 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 claims description 8
- 239000004471 Glycine Substances 0.000 claims description 8
- AEMRFAOFKBGASW-UHFFFAOYSA-N Glycolic acid Chemical compound OCC(O)=O AEMRFAOFKBGASW-UHFFFAOYSA-N 0.000 claims description 8
- CKLJMWTZIZZHCS-REOHCLBHSA-N L-aspartic acid Chemical compound OC(=O)[C@@H](N)CC(O)=O CKLJMWTZIZZHCS-REOHCLBHSA-N 0.000 claims description 8
- MTCFGRXMJLQNBG-UHFFFAOYSA-N Serine Natural products OCC(N)C(O)=O MTCFGRXMJLQNBG-UHFFFAOYSA-N 0.000 claims description 8
- 235000003704 aspartic acid Nutrition 0.000 claims description 8
- OQFSQFPPLPISGP-UHFFFAOYSA-N beta-carboxyaspartic acid Natural products OC(=O)C(N)C(C(O)=O)C(O)=O OQFSQFPPLPISGP-UHFFFAOYSA-N 0.000 claims description 8
- 239000008103 glucose Substances 0.000 claims description 8
- 229940054269 sodium pyruvate Drugs 0.000 claims description 8
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 claims description 7
- 235000012000 cholesterol Nutrition 0.000 claims description 7
- BVTBRVFYZUCAKH-UHFFFAOYSA-L disodium selenite Chemical compound [Na+].[Na+].[O-][Se]([O-])=O BVTBRVFYZUCAKH-UHFFFAOYSA-L 0.000 claims description 7
- 238000000605 extraction Methods 0.000 claims description 7
- 238000002844 melting Methods 0.000 claims description 7
- 229960001471 sodium selenite Drugs 0.000 claims description 7
- 239000011781 sodium selenite Substances 0.000 claims description 7
- 235000015921 sodium selenite Nutrition 0.000 claims description 7
- 239000008215 water for injection Substances 0.000 claims description 6
- 239000006285 cell suspension Substances 0.000 claims description 5
- 239000012228 culture supernatant Substances 0.000 claims description 5
- 238000001914 filtration Methods 0.000 claims description 5
- 239000002504 physiological saline solution Substances 0.000 claims description 5
- 238000010257 thawing Methods 0.000 claims description 5
- 238000005406 washing Methods 0.000 claims description 5
- ZSJLQEPLLKMAKR-UHFFFAOYSA-N Streptozotocin Natural products O=NN(C)C(=O)NC1C(O)OC(CO)C(O)C1O ZSJLQEPLLKMAKR-UHFFFAOYSA-N 0.000 claims description 3
- ZSJLQEPLLKMAKR-GKHCUFPYSA-N streptozocin Chemical compound O=NN(C)C(=O)N[C@H]1[C@@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O ZSJLQEPLLKMAKR-GKHCUFPYSA-N 0.000 claims description 3
- 229960001052 streptozocin Drugs 0.000 claims description 3
- 108010065805 Interleukin-12 Proteins 0.000 claims 1
- 102000013462 Interleukin-12 Human genes 0.000 claims 1
- 210000001519 tissue Anatomy 0.000 description 13
- 210000000130 stem cell Anatomy 0.000 description 9
- 239000006143 cell culture medium Substances 0.000 description 8
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 description 5
- 229930195725 Mannitol Natural products 0.000 description 5
- 230000000052 comparative effect Effects 0.000 description 5
- 239000000594 mannitol Substances 0.000 description 5
- 235000010355 mannitol Nutrition 0.000 description 5
- 238000012360 testing method Methods 0.000 description 5
- 108010023197 Streptokinase Proteins 0.000 description 4
- 238000000889 atomisation Methods 0.000 description 4
- 239000007640 basal medium Substances 0.000 description 4
- 229960005202 streptokinase Drugs 0.000 description 4
- 102100024222 B-lymphocyte antigen CD19 Human genes 0.000 description 3
- 102100037241 Endoglin Human genes 0.000 description 3
- 102000006354 HLA-DR Antigens Human genes 0.000 description 3
- 108010058597 HLA-DR Antigens Proteins 0.000 description 3
- 102100031573 Hematopoietic progenitor cell antigen CD34 Human genes 0.000 description 3
- 101000980825 Homo sapiens B-lymphocyte antigen CD19 Proteins 0.000 description 3
- 101000881679 Homo sapiens Endoglin Proteins 0.000 description 3
- 101000777663 Homo sapiens Hematopoietic progenitor cell antigen CD34 Proteins 0.000 description 3
- 101000946889 Homo sapiens Monocyte differentiation antigen CD14 Proteins 0.000 description 3
- 101000738771 Homo sapiens Receptor-type tyrosine-protein phosphatase C Proteins 0.000 description 3
- 101000800116 Homo sapiens Thy-1 membrane glycoprotein Proteins 0.000 description 3
- 241000598436 Human T-cell lymphotropic virus Species 0.000 description 3
- 102100035877 Monocyte differentiation antigen CD14 Human genes 0.000 description 3
- 241000204031 Mycoplasma Species 0.000 description 3
- 102100037422 Receptor-type tyrosine-protein phosphatase C Human genes 0.000 description 3
- 102100033523 Thy-1 membrane glycoprotein Human genes 0.000 description 3
- 241000589884 Treponema pallidum Species 0.000 description 3
- 238000004113 cell culture Methods 0.000 description 3
- 239000003814 drug Substances 0.000 description 3
- 239000002158 endotoxin Substances 0.000 description 3
- 239000003102 growth factor Substances 0.000 description 3
- 230000035755 proliferation Effects 0.000 description 3
- 210000002966 serum Anatomy 0.000 description 3
- MTCFGRXMJLQNBG-REOHCLBHSA-N (2S)-2-Amino-3-hydroxypropansäure Chemical compound OC[C@H](N)C(O)=O MTCFGRXMJLQNBG-REOHCLBHSA-N 0.000 description 2
- 206010011224 Cough Diseases 0.000 description 2
- 102000051325 Glucagon Human genes 0.000 description 2
- 108060003199 Glucagon Proteins 0.000 description 2
- 206010061218 Inflammation Diseases 0.000 description 2
- 229960005261 aspartic acid Drugs 0.000 description 2
- 230000010261 cell growth Effects 0.000 description 2
- 239000013592 cell lysate Substances 0.000 description 2
- 229940107161 cholesterol Drugs 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 208000035475 disorder Diseases 0.000 description 2
- MASNOZXLGMXCHN-ZLPAWPGGSA-N glucagon Chemical compound C([C@@H](C(=O)N[C@H](C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O)C(C)C)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@H](C)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](CO)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CCCCN)NC(=O)[C@H](CO)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](NC(=O)[C@H](CC=1C=CC=CC=1)NC(=O)[C@@H](NC(=O)CNC(=O)[C@H](CCC(N)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](N)CC=1NC=NC=1)[C@@H](C)O)[C@@H](C)O)C1=CC=CC=C1 MASNOZXLGMXCHN-ZLPAWPGGSA-N 0.000 description 2
- 229960004666 glucagon Drugs 0.000 description 2
- 229960002449 glycine Drugs 0.000 description 2
- 230000004054 inflammatory process Effects 0.000 description 2
- 210000004072 lung Anatomy 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 230000008439 repair process Effects 0.000 description 2
- 230000029058 respiratory gaseous exchange Effects 0.000 description 2
- 230000028327 secretion Effects 0.000 description 2
- 229960001153 serine Drugs 0.000 description 2
- 235000004400 serine Nutrition 0.000 description 2
- 208000025721 COVID-19 Diseases 0.000 description 1
- 206010008479 Chest Pain Diseases 0.000 description 1
- 102000008186 Collagen Human genes 0.000 description 1
- 108010035532 Collagen Proteins 0.000 description 1
- 201000004624 Dermatitis Diseases 0.000 description 1
- 238000008157 ELISA kit Methods 0.000 description 1
- RSEPBGGWRJCQGY-RBRWEJTLSA-N Estradiol valerate Chemical compound C1CC2=CC(O)=CC=C2[C@@H]2[C@@H]1[C@@H]1CC[C@H](OC(=O)CCCC)[C@@]1(C)CC2 RSEPBGGWRJCQGY-RBRWEJTLSA-N 0.000 description 1
- AEMRFAOFKBGASW-UHFFFAOYSA-M Glycolate Chemical compound OCC([O-])=O AEMRFAOFKBGASW-UHFFFAOYSA-M 0.000 description 1
- 206010068319 Oropharyngeal pain Diseases 0.000 description 1
- 206010037660 Pyrexia Diseases 0.000 description 1
- 239000000654 additive Substances 0.000 description 1
- 210000001789 adipocyte Anatomy 0.000 description 1
- 230000032683 aging Effects 0.000 description 1
- 230000003110 anti-inflammatory effect Effects 0.000 description 1
- 230000001754 anti-pyretic effect Effects 0.000 description 1
- 239000002221 antipyretic Substances 0.000 description 1
- 230000006907 apoptotic process Effects 0.000 description 1
- 208000010668 atopic eczema Diseases 0.000 description 1
- 230000009286 beneficial effect Effects 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
- 230000015572 biosynthetic process Effects 0.000 description 1
- 230000021164 cell adhesion Effects 0.000 description 1
- 230000004663 cell proliferation Effects 0.000 description 1
- 230000003833 cell viability Effects 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 210000001612 chondrocyte Anatomy 0.000 description 1
- 229920001436 collagen Polymers 0.000 description 1
- 238000013329 compounding Methods 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 235000014113 dietary fatty acids Nutrition 0.000 description 1
- 230000004069 differentiation Effects 0.000 description 1
- 230000003467 diminishing effect Effects 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 229960004766 estradiol valerate Drugs 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 229930195729 fatty acid Natural products 0.000 description 1
- 239000000194 fatty acid Substances 0.000 description 1
- 150000004665 fatty acids Chemical class 0.000 description 1
- 229960001031 glucose Drugs 0.000 description 1
- 229960004275 glycolic acid Drugs 0.000 description 1
- 230000001339 gustatory effect Effects 0.000 description 1
- 230000002519 immonomodulatory effect Effects 0.000 description 1
- 210000000987 immune system Anatomy 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 210000003716 mesoderm Anatomy 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 210000002894 multi-fate stem cell Anatomy 0.000 description 1
- 210000000663 muscle cell Anatomy 0.000 description 1
- 238000002663 nebulization Methods 0.000 description 1
- 210000000653 nervous system Anatomy 0.000 description 1
- 210000002569 neuron Anatomy 0.000 description 1
- 235000021048 nutrient requirements Nutrition 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 210000000963 osteoblast Anatomy 0.000 description 1
- 230000002611 ovarian Effects 0.000 description 1
- 230000035790 physiological processes and functions Effects 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 235000018102 proteins Nutrition 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 208000023409 throat pain Diseases 0.000 description 1
- 230000017423 tissue regeneration Effects 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
- 150000003722 vitamin derivatives Chemical class 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- A61K38/18—Growth factors; Growth regulators
- A61K38/1825—Fibroblast growth factor [FGF]
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- A61K38/18—Growth factors; Growth regulators
- A61K38/1808—Epidermal growth factor [EGF] urogastrone
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- A61K38/18—Growth factors; Growth regulators
- A61K38/1833—Hepatocyte growth factor; Scatter factor; Tumor cytotoxic factor II
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- A61K38/18—Growth factors; Growth regulators
- A61K38/1858—Platelet-derived growth factor [PDGF]
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- A61K38/18—Growth factors; Growth regulators
- A61K38/1858—Platelet-derived growth factor [PDGF]
- A61K38/1866—Vascular endothelial growth factor [VEGF]
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- A61K38/19—Cytokines; Lymphokines; Interferons
- A61K38/191—Tumor necrosis factors [TNF], e.g. lymphotoxin [LT], i.e. TNF-beta
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- A61K38/19—Cytokines; Lymphokines; Interferons
- A61K38/193—Colony stimulating factors [CSF]
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- A61K38/19—Cytokines; Lymphokines; Interferons
- A61K38/195—Chemokines, e.g. RANTES
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- A61K38/19—Cytokines; Lymphokines; Interferons
- A61K38/20—Interleukins [IL]
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- A61K38/19—Cytokines; Lymphokines; Interferons
- A61K38/20—Interleukins [IL]
- A61K38/2006—IL-1
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- A61K38/19—Cytokines; Lymphokines; Interferons
- A61K38/20—Interleukins [IL]
- A61K38/2013—IL-2
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- A61K38/19—Cytokines; Lymphokines; Interferons
- A61K38/20—Interleukins [IL]
- A61K38/2026—IL-4
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- A61K38/19—Cytokines; Lymphokines; Interferons
- A61K38/20—Interleukins [IL]
- A61K38/2033—IL-5
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- A61K38/19—Cytokines; Lymphokines; Interferons
- A61K38/20—Interleukins [IL]
- A61K38/204—IL-6
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- A61K38/19—Cytokines; Lymphokines; Interferons
- A61K38/20—Interleukins [IL]
- A61K38/2046—IL-7
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- A61K38/19—Cytokines; Lymphokines; Interferons
- A61K38/20—Interleukins [IL]
- A61K38/2053—IL-8
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- A61K38/19—Cytokines; Lymphokines; Interferons
- A61K38/20—Interleukins [IL]
- A61K38/206—IL-9
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- A61K38/19—Cytokines; Lymphokines; Interferons
- A61K38/20—Interleukins [IL]
- A61K38/2066—IL-10
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- A61K38/19—Cytokines; Lymphokines; Interferons
- A61K38/20—Interleukins [IL]
- A61K38/208—IL-12
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- A61K38/19—Cytokines; Lymphokines; Interferons
- A61K38/20—Interleukins [IL]
- A61K38/2086—IL-13 to IL-16
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- A61K38/19—Cytokines; Lymphokines; Interferons
- A61K38/21—Interferons [IFN]
- A61K38/217—IFN-gamma
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/475—Growth factors; Growth regulators
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/475—Growth factors; Growth regulators
- C07K14/4753—Hepatocyte growth factor; Scatter factor; Tumor cytotoxic factor II
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/475—Growth factors; Growth regulators
- C07K14/485—Epidermal growth factor [EGF], i.e. urogastrone
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/475—Growth factors; Growth regulators
- C07K14/49—Platelet-derived growth factor [PDGF]
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/475—Growth factors; Growth regulators
- C07K14/50—Fibroblast growth factor [FGF]
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/52—Cytokines; Lymphokines; Interferons
- C07K14/521—Chemokines
- C07K14/522—Alpha-chemokines, e.g. NAP-2, ENA-78, GRO-alpha/MGSA/NAP-3, GRO-beta/MIP-2alpha, GRO-gamma/MIP-2beta, IP-10, GCP-2, MIG, PBSF, PF-4, KC
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/52—Cytokines; Lymphokines; Interferons
- C07K14/521—Chemokines
- C07K14/523—Beta-chemokines, e.g. RANTES, I-309/TCA-3, MIP-1alpha, MIP-1beta/ACT-2/LD78/SCIF, MCP-1/MCAF, MCP-2, MCP-3, LDCF-1, LDCF-2
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/52—Cytokines; Lymphokines; Interferons
- C07K14/525—Tumour necrosis factor [TNF]
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/52—Cytokines; Lymphokines; Interferons
- C07K14/53—Colony-stimulating factor [CSF]
- C07K14/535—Granulocyte CSF; Granulocyte-macrophage CSF
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/52—Cytokines; Lymphokines; Interferons
- C07K14/54—Interleukins [IL]
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/52—Cytokines; Lymphokines; Interferons
- C07K14/54—Interleukins [IL]
- C07K14/5406—IL-4
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/52—Cytokines; Lymphokines; Interferons
- C07K14/54—Interleukins [IL]
- C07K14/5409—IL-5
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/52—Cytokines; Lymphokines; Interferons
- C07K14/54—Interleukins [IL]
- C07K14/5412—IL-6
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/52—Cytokines; Lymphokines; Interferons
- C07K14/54—Interleukins [IL]
- C07K14/5418—IL-7
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/52—Cytokines; Lymphokines; Interferons
- C07K14/54—Interleukins [IL]
- C07K14/5421—IL-8
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/52—Cytokines; Lymphokines; Interferons
- C07K14/54—Interleukins [IL]
- C07K14/5425—IL-9
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/52—Cytokines; Lymphokines; Interferons
- C07K14/54—Interleukins [IL]
- C07K14/5428—IL-10
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/52—Cytokines; Lymphokines; Interferons
- C07K14/54—Interleukins [IL]
- C07K14/5434—IL-12
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/52—Cytokines; Lymphokines; Interferons
- C07K14/54—Interleukins [IL]
- C07K14/5437—IL-13
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/52—Cytokines; Lymphokines; Interferons
- C07K14/54—Interleukins [IL]
- C07K14/5443—IL-15
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/52—Cytokines; Lymphokines; Interferons
- C07K14/54—Interleukins [IL]
- C07K14/545—IL-1
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/52—Cytokines; Lymphokines; Interferons
- C07K14/54—Interleukins [IL]
- C07K14/55—IL-2
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/52—Cytokines; Lymphokines; Interferons
- C07K14/555—Interferons [IFN]
- C07K14/57—IFN-gamma
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/06—Animal cells or tissues; Human cells or tissues
- C12N5/0602—Vertebrate cells
- C12N5/0652—Cells of skeletal and connective tissues; Mesenchyme
- C12N5/0662—Stem cells
- C12N5/0668—Mesenchymal stem cells from other natural sources
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P21/00—Preparation of peptides or proteins
- C12P21/02—Preparation of peptides or proteins having a known sequence of two or more amino acids, e.g. glutathione
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2500/00—Specific components of cell culture medium
- C12N2500/30—Organic components
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2500/00—Specific components of cell culture medium
- C12N2500/30—Organic components
- C12N2500/32—Amino acids
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2500/00—Specific components of cell culture medium
- C12N2500/30—Organic components
- C12N2500/34—Sugars
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2500/00—Specific components of cell culture medium
- C12N2500/30—Organic components
- C12N2500/36—Lipids
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
- C12N2501/30—Hormones
- C12N2501/33—Insulin
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
- C12N2501/30—Hormones
- C12N2501/335—Glucagon; Glucagon-like peptide [GLP]; Exendin
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
- C12N2501/70—Enzymes
- C12N2501/73—Hydrolases (EC 3.)
- C12N2501/734—Proteases (EC 3.4.)
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
- C12N2501/998—Proteins not provided for elsewhere
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Gastroenterology & Hepatology (AREA)
- General Health & Medical Sciences (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Medicinal Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Genetics & Genomics (AREA)
- Biochemistry (AREA)
- Molecular Biology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biophysics (AREA)
- Toxicology (AREA)
- Immunology (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Animal Behavior & Ethology (AREA)
- Epidemiology (AREA)
- Pharmacology & Pharmacy (AREA)
- Wood Science & Technology (AREA)
- Biotechnology (AREA)
- Biomedical Technology (AREA)
- General Engineering & Computer Science (AREA)
- Developmental Biology & Embryology (AREA)
- Microbiology (AREA)
- Vascular Medicine (AREA)
- General Chemical & Material Sciences (AREA)
- Rheumatology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Cell Biology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
本发明提供一种具有改善呼吸系统感染作用的全阵列细胞因子的制备方法，全阵列细胞因子包括FGF、G‑CSF、GM‑CSF、IFN‑γ、IL‑1β、IL‑1ra、IL‑2、IL‑4、IL‑5、IL‑6、IL‑7、IL‑8、IL‑9、IL‑10、IL‑12p70、IL‑13、IL‑15、IL‑17A、IP‑10、MCP‑1/MCAF、MIP‑1α、MIP‑1β、PDGF‑BB、RANTES、TNF‑α、VEGF、EGF、HGF，本发明制得的全阵列细胞因子具有良好的改善呼吸系统感染的作用。
Description
技术领域
本发明涉及细胞因子提取技术领域，特别涉及一种具有改善呼吸系统感染作用的全阵列细胞因子及其制备方法。
背景技术
间充质干细胞(MSC)是属于中胚层的，具有高度自我更新能力和多向分化潜能的多能干细胞。广泛存在于全身多种组织中，可在体外培养扩增，并能在特定条件下分化为神经细胞、成骨细胞、软骨细胞、肌肉细胞、脂肪细胞等。此外，间充质干细胞还具有强大的分泌能力，其分泌的细胞因子可以促进其它细胞增殖、修复受损组织、调节免疫系统、神经系统、抑制炎症等。正因如此，干细胞产生的生长因子被广泛应用于组织再生、免疫调节等领域。由于脐带中含有大量的间充质干细胞，且采集过程不会对供体造成任何损伤，所以脐带间充质干细胞的生长因子被广泛使用。例如专利CN113662967A提供了一种脐带干细胞裂解液治疗卵巢衰老方法；专利CN110448682A提供了一种使用干细胞裂解液治疗湿疹的方法等。本专利详细阐述了一种脐带间充质干细胞全阵列细胞因子的提取方法，以及使用脐带间充质干细胞全阵列细胞因子治疗COVID-19引起的呼吸系统感染的方法。
发明内容
鉴于此，本发明提出一种具有改善呼吸系统感染作用的全阵列细胞因子的制备方法，解决上述问题。
本发明的技术方案是这样实现的：
一种具有改善呼吸系统感染作用的全阵列细胞因子，全阵列细胞因子包括FGF、G-CSF、GM-CSF、IFN-γ、IL-1β、IL-1ra、IL-2、IL-4、IL-5、IL-6、IL-7、IL-8、IL-9、IL-10、IL-12p70、IL-13、IL-15、IL-17A、IP-10、MCP-1/MCAF、MIP-1α、MIP-1β、PDGF-BB、RANTES、TNF-α、VEGF、EGF、HGF。
一种具有改善呼吸系统感染作用的全阵列细胞因子的制备方法，包括以下步骤：
(1)脐带组织采用组织块贴壁法分离培养脐带间充质干细胞，得到原代细胞；
(2)细胞传代：原代细胞汇合度达到80％～90％时传代，收集第5代细胞培养上清液，冻干，得到传代细胞；
(3)细胞复融：将传代细胞在37℃水浴复融，细胞悬液用10倍体积的生理盐水洗涤一次后计数，离心，弃上清，得到复融后的细胞；
(4)细胞因子提取：将复融后的细胞采用注射用水重悬，将重悬细胞在23-27℃静置10-15min，再将重悬细胞浸入液氮中，得到冻结后的重悬细胞，冻结后的重悬细胞放入水浴锅中复融，复融后的重悬细胞浸入液氮中，重复冻融2-5次，收集上清液，过滤，制得全阵列细胞因子。
进一步的，步骤(2)中，细胞传代培养基包括基础培养基、纤粘连蛋白、胰岛素、丙酮酸钠、葡萄糖和甘醇酸。
进一步的，所述基础培养基为DMEM培养基。
进一步的，所述纤粘连蛋白8-12μg/mL、胰岛素13-17ng/mL、丙酮酸钠180-200mg/L、葡萄糖2-4g/L。
进一步的，所述甘醇酸浓度为13-17μM。
本发明传代培养基中组分合理配比，使原代细胞能够快速增殖分裂，能够诱导原代细胞分泌目标细胞因子，本发明传代培养基中甘醇酸不仅能加快细胞的增殖还能够增加细胞的黏着性，避免细胞产生机械损伤，本发明获得的第五代细胞细胞活力更好，稳定性更高。
进一步的，步骤(1)中，将脐带组织剪至1-3mm3，将剪碎后的脐带组织加入传代培养基中进行培养。
进一步的，所述传代培养基包括基础培养基、胰岛素、胰高糖素、链丝菌蛋白酶、亚硒酸钠、天门冬氨酸、丝氨酸、甘氨酸、胆固醇和纤粘连蛋白。
进一步的，所述基础培养基为SFM培养基，胰岛素25-35μg/mL、链丝菌蛋白酶0.4-0.6g/L、亚硒酸钠13-17μg/mL、天门冬氨酸7.5mg/L～11.5mg、丝氨酸8mg/L～14mg/L、甘氨酸6mg/L～8mg/L、胆固醇25-45mg/L和纤粘连蛋白23-27μg/mL。
本发明提取原代细胞培养基采用SFM培养基、胰岛素、胰高糖素、链丝菌蛋白酶、亚硒酸钠、天门冬氨酸、丝氨酸、甘氨酸、胆固醇和纤粘连蛋白制得。由于本发明提取原料为脐带组织，因此本发明原代细胞培养基不涉及血清添加物，可以减少血清中未知组分的干扰和血源性污染，减少血清的添加会导致细胞增长过慢、容易受到机械和化学因素的损伤、细胞增殖过慢等问题出现。本发明中通过SFM培养基和补充因子混合，能够促进细胞RNA、蛋白质和脂肪酸合成，防止细胞凋亡。合理的营养成分能够满足细胞生长的营养需求，提高目的产物的表达量。
进一步的，全阵列细胞因子冻干剂采用雾化给药的方式进行给药。
进一步的，根据全阵列细胞因子加入0.1％的甘露醇进行冻干。
进一步的，冻干参数为在-30℃下预冻5h，调整冷冻干燥机温度-60℃，真空度为3Pa下冻干10h。
与现有技术相比，本发明的有益效果是：
本发明采用干细胞全阵列因子作为雾化药物，因其含有丰富的生长因子、抗炎因子，可快速起到消炎止咳的作用，并且促进肺部及呼吸道细胞更新，快速修复肺部及呼吸道生理功能。
具体实施方式
为了更好理解本发明技术内容，下面提供具体实施例，对本发明做进一步的说明。
本发明实施例所用的实验方法如无特殊说明，均为常规方法。
本发明实施例所用的材料、试剂等，如无特殊说明，均可从商业途径得到。
实施例1
原代细胞培养基由SFM培养基、胰岛素25μg/mL、链丝菌蛋白酶0.4g/L、亚硒酸钠13μg/mL、天门冬氨酸7.511.5mg、丝氨酸8mg/L、甘氨酸6mg/L、胆固醇25mg/L和纤粘连蛋白23μg/mL组成。
细胞传代培养基由DMEM培养基、纤粘连蛋白8μg/mL、胰岛素13ng/mL、丙酮酸钠180mg/L、葡萄糖2g/L、甘醇酸浓度为13μM组成。
全阵列细胞因子的制备方法，由以下步骤制得：
(1)将脐带组织剪至2mm3，将剪碎后的脐带组织加入原代培养基中，于37℃，5％CO2培养箱中进行培养，汇合度达到80％时，送检HBV、HIV、HCV、梅毒螺旋体、HTLV、CMV、EBV，检测符合质量要求的样本，得到原代细胞；
(2)细胞传代：将原代细胞，细胞密度为10000个细胞/cm2，放进细胞传代培养基中培养收集第5代细胞培养上清液，在37℃、5％CO2培养箱中进行培养，待细胞汇合度达到80％～90％时冻存细胞，并送检无菌、支原体、内毒素和细胞表型(CD73、CD90、CD105、CD45、CD34、CD19、CD14、HLA-DR等)，符合质量要求的细胞可用于细胞因子提取；
(3)细胞复融：从液氮罐中取出细胞，在37℃水浴中快速复融。细胞悬液用10倍体积的生理盐水洗涤一次后，离心，弃上清；
(4)细胞因子提取：将复融后的细胞采用注射用水重悬，将重悬细胞在23℃静置15min，再将重悬细胞浸入液氮中，得到冻结后的重悬细胞，冻结后的重悬细胞放入37℃水浴锅中复融，复融后的重悬细胞浸入液氮中，重复冻融4次，收集上清液，过滤，制得全阵列细胞因子，根据全阵列细胞因子体积加入0.1％的甘露醇进行冻干，在-30℃下预冻5h，调整冷冻干燥机温度-60℃，真空度为3Pa下冻干10h。
实施例2
原代细胞培养基由SFM培养基、胰岛素35μg/mL、链丝菌蛋白酶0.6g/L、亚硒酸钠17μg/mL、天门冬氨酸11.5mg、丝氨酸14mg/L、甘氨酸8mg/L、胆固醇45mg/L和纤粘连蛋白27μg/mL组成。
细胞传代培养基由DMEM培养基、纤粘连蛋白12μg/mL、胰岛素17ng/mL、丙酮酸钠200mg/L、葡萄糖4g/L、甘醇酸浓度为17μM组成。
全阵列细胞因子的制备方法，由以下步骤制得：
(1)将脐带组织剪至2mm3，将剪碎后的脐带组织加入原代培养基中，于37℃，5％CO2培养箱中进行培养，汇合度达到90％时，送检HBV、HIV、HCV、梅毒螺旋体、HTLV、CMV、EBV，检测符合质量要求的样本，得到原代细胞；
(2)将原代细胞，细胞密度为10000个细胞/cm2，放进细胞传代培养基中培养收集第5代细胞培养上清液，在37℃、5％CO2培养箱中进行培养，待细胞汇合度达到80％～90％时冻存细胞，并送检无菌、支原体、内毒素和细胞表型(CD73、CD90、CD105、CD45、CD34、CD19、CD14、HLA-DR等)，符合质量要求的细胞可用于细胞因子提取；
(3)细胞复融：从液氮罐中取出细胞，在37℃水浴中快速复融。细胞悬液用10倍体积的生理盐水洗涤一次后，离心，弃上清；
(4)细胞因子提取：将复融后的细胞采用注射用水重悬，将重悬细胞在23℃静置15min，再将重悬细胞浸入液氮中，得到冻结后的重悬细胞，冻结后的重悬细胞放入37℃水浴锅中复融，复融后的重悬细胞浸入液氮中，重复冻融4次，收集上清液，过滤，制得全阵列细胞因子，根据全阵列细胞因子体积加入0.1％的甘露醇进行冻干，在-30℃下预冻5h，调整冷冻干燥机温度-60℃，真空度为3Pa下冻干10h。
实施例3
原代细胞培养基由SFM培养基、胰岛素30μg/mL、链丝菌蛋白酶0.5g/L、亚硒酸钠15μg/mL、天门冬氨酸9mg、丝氨酸11mg/L、甘氨酸7mg/L、胆固醇35mg/L和纤粘连蛋白24μg/mL组成。
细胞传代培养基由DMEM培养基、纤粘连蛋白10μg/mL、胰岛素15ng/mL、丙酮酸钠190mg/L、葡萄糖3g/L、甘醇酸浓度为15μM组成。
全阵列细胞因子的制备方法，由以下步骤制得：
(1)将脐带组织剪至1-3mm3，将剪碎后的脐带组织加入原代培养基中，于37℃，5％CO2培养箱中进行培养，汇合度达到85％时，送检HBV、HIV、HCV、梅毒螺旋体、HTLV、CMV、EBV，检测符合质量要求的样本，得到原代细胞；
(2)细胞传代：将原代细胞，细胞密度为10000个细胞/cm2，放进细胞传代培养基中培养收集第5代细胞培养上清液，在37℃、5％CO2培养箱中进行培养，待细胞汇合度达到80％～90％时冻存细胞，并送检无菌、支原体、内毒素和细胞表型(CD73、CD90、CD105、CD45、CD34、CD19、CD14、HLA-DR等)，符合质量要求的细胞可用于细胞因子提取；
(3)细胞复融：从液氮罐中取出细胞，在37℃水浴中快速复融。细胞悬液用10倍体积的生理盐水洗涤一次后，离心，弃上清；
(4)细胞因子提取：将复融后的细胞采用注射用水重悬，将重悬细胞在23℃静置15min，再将重悬细胞浸入液氮中，得到冻结后的重悬细胞，冻结后的重悬细胞放入37℃水浴锅中复融，复融后的重悬细胞浸入液氮中，重复冻融4次，收集上清液，过滤，制得全阵列细胞因子，根据全阵列细胞因子体积加入0.1％的甘露醇进行冻干，在-30℃下预冻5h，调整冷冻干燥机温度-60℃，真空度为3Pa下冻干10h。
对比例1
在实施例3的基础上调整原代细胞培养基的组成，具体为：SFM培养基、胰岛素30μg/mL、戊酸雌二醇10ng/mL、天门冬氨酸9mg、丝氨酸11mg/L、甘氨酸7mg/L和纤粘连蛋白24μg/mL组成。
对比例2
在实施例3的基础上调整细胞传代培养基的组成，具体为：细胞传代培养基由DMEM培养基、胶原蛋白10μg/mL、胰岛素15ng/mL、维生素B2 0.3mg/L、丙酮酸钠190mg/L和葡萄糖3g/L组成。
对比例3
在实施例3的基础上调整细胞传代培养基的配比，具体为：细胞传代培养基由DMEM培养基、纤粘连蛋白16μg/mL、胰岛素23ng/mL、丙酮酸钠160mg/L、葡萄糖6g/L、甘醇酸浓度为15μM组成。
试验例1
采用实施例3制得干细胞全阵列因子作为雾化药物，将冻干后的干细胞全阵列因子5mg加30ml的注射用水溶解，采用映美HN300雾化器进行雾化，雾化时间为30min，试验地点为海南一龄医疗产业发展有限公司，时间为2022年12月，受试者共50位，其中有嗅觉障碍/味觉障碍的2人、活动后呼吸急促的3人、持续咳嗽45人、胸痛27人、呼吸21人和咽喉疼痛33人。在治疗期间，如果患者发烧，则需要服用退烧药，此外，患者未使用任何其它药物。每天每位患者接受一次雾化治疗，时间在30分钟左右，具体疗效见下表：
试验例2
采用ELISA试剂盒检测实施例3和对比例1-3制得的全阵列细胞因子中EGF、FGF、VEGF的含量。
试验结果表明，本发明中原代细胞培养基和细胞传代培养基能够促进细胞因子分泌增殖。
试验例3
将施例3和对比例1-3制得的全阵列细胞因子冻干前和冻干后细胞存活率测试，采用细胞荧光分析仪进行采集数据。
实验结果表明，本发明制得的干细胞全阵列因子和甘露醇复配冻干可以得到很好的保存效果，进一步表明本发明中原代细胞培养基和细胞传代培养基能够提高细胞的活力和稳定性。
以上所述仅为本发明的较佳实施例而已，并不用以限制本发明，凡在本发明的精神和原则之内，所作的任何修改、等同替换、改进等，均应包含在本发明的保护范围之内。
Claims (1)
1.一种具有改善呼吸系统感染作用的全阵列细胞因子的制备方法，其特征在于：包括以下步骤：
（1）将脐带组织剪至1-3mm3，将剪碎后的脐带组织加入原代培养基中进行培养，脐带组织采用组织块贴壁法分离培养脐带间充质干细胞，得到原代细胞；所述原代培养基包括SFM培养基、胰岛素25-35μg/mL、链丝菌蛋白酶0.4-0.6g/L、亚硒酸钠13-17μg/mL、天门冬氨酸7.5～11.5mg、丝氨酸8～14mg/L、甘氨酸6～8mg/L、胆固醇25-45mg/L和纤粘连蛋白23-27μg/mL；
（2）细胞传代：原代细胞汇合度达到80%～90%时放进细胞传代培养基中培养，收集第5代细胞培养上清液，冻干，得到传代细胞；所述细胞传代培养基包括DMEM培养基、纤粘连蛋白8-12μg/mL、胰岛素13-17ng/mL、丙酮酸钠180-200mg/L、葡萄糖2-4g/L、甘醇酸浓度为13-17μM；
（3）细胞复融：将传代细胞水浴复融，细胞悬液用8-12倍体积的生理盐水洗涤一次后计数，离心，弃上清，得到复融后的细胞；
细胞因子提取：将复融后的细胞采用注射用水重悬，将重悬细胞在23-27℃静置10-15min，再将重悬细胞浸入液氮中，得到冻结后的重悬细胞，冻结后的重悬细胞放入水浴锅中复融，复融后的重悬细胞浸入液氮中，重复冻融2-5次，收集上清液，过滤，制得全阵列细胞因子；
所述全阵列细胞因子包括：FGF、G-CSF、GM-CSF、IFN-γ、IL-1β、IL-1ra、IL-2、IL-4、IL-5、IL-6、IL-7、IL-8、IL-9、IL-10、IL-12 p70、IL-13、IL-15、IL-17A、IP-10、MCP-1/MCAF、MIP-1α、MIP-1β、PDGF-BB、RANTES、TNF-α、VEGF、EGF、HGF。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310630711.6A CN117122668B (zh) | 2023-05-31 | 2023-05-31 | 一种具有改善呼吸系统感染作用的全阵列细胞因子及其制备方法 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310630711.6A CN117122668B (zh) | 2023-05-31 | 2023-05-31 | 一种具有改善呼吸系统感染作用的全阵列细胞因子及其制备方法 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN117122668A CN117122668A (zh) | 2023-11-28 |
| CN117122668B true CN117122668B (zh) | 2024-06-11 |
Family
ID=88853373
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202310630711.6A Active CN117122668B (zh) | 2023-05-31 | 2023-05-31 | 一种具有改善呼吸系统感染作用的全阵列细胞因子及其制备方法 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN117122668B (zh) |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106520689A (zh) * | 2016-12-05 | 2017-03-22 | 四川华皓生物科技有限公司 | 一种间充质干细胞细胞因子的制备方法和应用 |
| WO2017096618A1 (zh) * | 2015-12-11 | 2017-06-15 | 郭镭 | 一种从脐带外层羊膜组织中分离培养脐带间充质干细胞的方法 |
| CN110403958A (zh) * | 2019-08-08 | 2019-11-05 | 辰信(上海)医疗科技有限公司 | 干细胞活性肽雾化吸入法治疗肺纤维化及雾化剂的备制 |
| CN111394304A (zh) * | 2020-03-27 | 2020-07-10 | 北京臻溪谷医学研究中心（有限合伙） | 一种修复呼吸系统损伤的干细胞条件培养上清液及其制剂 |
-
2023
- 2023-05-31 CN CN202310630711.6A patent/CN117122668B/zh active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2017096618A1 (zh) * | 2015-12-11 | 2017-06-15 | 郭镭 | 一种从脐带外层羊膜组织中分离培养脐带间充质干细胞的方法 |
| CN106520689A (zh) * | 2016-12-05 | 2017-03-22 | 四川华皓生物科技有限公司 | 一种间充质干细胞细胞因子的制备方法和应用 |
| CN110403958A (zh) * | 2019-08-08 | 2019-11-05 | 辰信(上海)医疗科技有限公司 | 干细胞活性肽雾化吸入法治疗肺纤维化及雾化剂的备制 |
| CN111394304A (zh) * | 2020-03-27 | 2020-07-10 | 北京臻溪谷医学研究中心（有限合伙） | 一种修复呼吸系统损伤的干细胞条件培养上清液及其制剂 |
Non-Patent Citations (1)
| Title |
|---|
| Nebulization Therapy with Umbilical Cord Mesenchymal Stem Cell‑Derived Exosomes for COVID‑19 Pneumonia;Meiping Chu等;《Stem Cell Reviews and Reports》;20220604;第18卷;第2152-2163页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN117122668A (zh) | 2023-11-28 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103589683A (zh) | 一种脐带间充质干细胞的分离方法和培养方法 | |
| CN112725270B (zh) | 一种人源骨髓间充质干细胞诱导培养基及诱导方法 | |
| CN105663168A (zh) | 用于修复卵巢功能的细胞制剂 | |
| CN115025123B (zh) | 肝巨噬细胞调节剂及其制备方法和应用 | |
| CN101514333B (zh) | 一种免疫耐受型树突状细胞及其制备方法与专用培养基 | |
| CN111973632B (zh) | 一种用于治疗糖尿病的干细胞制剂及其制备方法 | |
| CN115300612A (zh) | 一种修复子宫内膜的干细胞制剂及其应用 | |
| CN102965337A (zh) | 分离提取人皮下脂肪间充质干细胞的方法和提取专用培养基 | |
| CN117122668B (zh) | 一种具有改善呼吸系统感染作用的全阵列细胞因子及其制备方法 | |
| CN103421740B (zh) | 一种人骨髓间充质干细胞体外培养扩增方法 | |
| CN114621916A (zh) | 抗氧化剂在提高脐带间充质干细胞活力中的应用 | |
| CN112300986B (zh) | 无血清培养基制备脂肪间充质干细胞的方法 | |
| CN114164171A (zh) | 一种人脐带间充质干细胞培养基、注射液、制备方法与其在制备治疗脑卒中的药物中的应用 | |
| CN114515334B (zh) | 一种脂肪间充质干细胞因子冻干粉及其制备方法 | |
| CN115305232B (zh) | 一种脂肪间充质干细胞复苏培养液及复苏方法 | |
| CN112708595B (zh) | 一种sd大鼠来源骨髓间充质干细胞诱导培养基及诱导方法 | |
| CN112877286B (zh) | 一种干细胞体外诱导方法 | |
| EP3563861A1 (en) | Ginseng-derived nanoparticle and preparation and application thereof | |
| CN109576308A (zh) | 一种提高人类干细胞来源肝样细胞解毒功能的方法及其应用 | |
| CN109749981B (zh) | 人源脂肪干细胞来源的肝细胞样细胞及其制备方法和应用 | |
| CN112522193A (zh) | 无血清培养基在分离和传代脂肪间充质干细胞中的用途 | |
| JP2011200180A (ja) | 幹細胞の未分化維持剤及び増殖促進剤 | |
| TWI715874B (zh) | 無血清培養液以及使用此無血清培養液分離和放大培養幹細胞方法 | |
| JPS63304978A (ja) | 上皮細胞の増殖法 | |
| WO2014075634A1 (zh) | 间质血管层细胞和间充质祖细胞在预防或治疗骨性关节炎中的应用 |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |