JPWO2018123968A1 - 生体組織損傷の修復剤および当該修復剤の製造方法 - Google Patents
生体組織損傷の修復剤および当該修復剤の製造方法 Download PDFInfo
- Publication number
- JPWO2018123968A1 JPWO2018123968A1 JP2018559456A JP2018559456A JPWO2018123968A1 JP WO2018123968 A1 JPWO2018123968 A1 JP WO2018123968A1 JP 2018559456 A JP2018559456 A JP 2018559456A JP 2018559456 A JP2018559456 A JP 2018559456A JP WO2018123968 A1 JPWO2018123968 A1 JP WO2018123968A1
- Authority
- JP
- Japan
- Prior art keywords
- mesenchymal stem
- stem cells
- serum
- repair agent
- cells
- 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.)
- Granted
Links
- 230000008439 repair process Effects 0.000 title claims abstract description 71
- 230000000451 tissue damage Effects 0.000 title claims abstract description 45
- 231100000827 tissue damage Toxicity 0.000 title claims abstract description 45
- 238000004519 manufacturing process Methods 0.000 title claims abstract description 28
- 210000002901 mesenchymal stem cell Anatomy 0.000 claims abstract description 204
- 239000012679 serum free medium Substances 0.000 claims abstract description 116
- 239000003795 chemical substances by application Substances 0.000 claims abstract description 101
- 210000004027 cell Anatomy 0.000 claims description 126
- 102100032807 Tumor necrosis factor-inducible gene 6 protein Human genes 0.000 claims description 61
- 101710169430 Tumor necrosis factor-inducible gene 6 protein Proteins 0.000 claims description 61
- 238000000034 method Methods 0.000 claims description 41
- 206010016654 Fibrosis Diseases 0.000 claims description 40
- 210000002540 macrophage Anatomy 0.000 claims description 38
- 230000000694 effects Effects 0.000 claims description 37
- 210000001519 tissue Anatomy 0.000 claims description 37
- 201000010099 disease Diseases 0.000 claims description 36
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims description 36
- 230000004761 fibrosis Effects 0.000 claims description 31
- 238000012258 culturing Methods 0.000 claims description 30
- 239000003814 drug Substances 0.000 claims description 29
- 229940079593 drug Drugs 0.000 claims description 27
- 150000003904 phospholipids Chemical class 0.000 claims description 27
- 235000014113 dietary fatty acids Nutrition 0.000 claims description 25
- 229930195729 fatty acid Natural products 0.000 claims description 25
- 239000000194 fatty acid Substances 0.000 claims description 25
- 210000000988 bone and bone Anatomy 0.000 claims description 24
- 150000004665 fatty acids Chemical class 0.000 claims description 23
- 210000004969 inflammatory cell Anatomy 0.000 claims description 18
- 230000008595 infiltration Effects 0.000 claims description 17
- 238000001764 infiltration Methods 0.000 claims description 17
- 208000020832 chronic kidney disease Diseases 0.000 claims description 14
- 208000019425 cirrhosis of liver Diseases 0.000 claims description 13
- 230000007547 defect Effects 0.000 claims description 11
- 206010019728 Hepatitis alcoholic Diseases 0.000 claims description 9
- 206010019799 Hepatitis viral Diseases 0.000 claims description 9
- 208000002353 alcoholic hepatitis Diseases 0.000 claims description 9
- 201000001862 viral hepatitis Diseases 0.000 claims description 9
- 206010028980 Neoplasm Diseases 0.000 claims description 8
- 201000006417 multiple sclerosis Diseases 0.000 claims description 8
- 208000024827 Alzheimer disease Diseases 0.000 claims description 7
- 206010003210 Arteriosclerosis Diseases 0.000 claims description 7
- 206010007710 Cartilage injury Diseases 0.000 claims description 7
- 208000006545 Chronic Obstructive Pulmonary Disease Diseases 0.000 claims description 7
- 206010009269 Cleft palate Diseases 0.000 claims description 7
- 208000003556 Dry Eye Syndromes Diseases 0.000 claims description 7
- 206010013774 Dry eye Diseases 0.000 claims description 7
- 208000029523 Interstitial Lung disease Diseases 0.000 claims description 7
- 241000124008 Mammalia Species 0.000 claims description 7
- 201000007100 Pharyngitis Diseases 0.000 claims description 7
- 206010035664 Pneumonia Diseases 0.000 claims description 7
- 206010037765 Radiation pneumonitis Diseases 0.000 claims description 7
- 208000006011 Stroke Diseases 0.000 claims description 7
- 208000011775 arteriosclerosis disease Diseases 0.000 claims description 7
- 206010003246 arthritis Diseases 0.000 claims description 7
- 239000000316 bone substitute Substances 0.000 claims description 7
- 201000011510 cancer Diseases 0.000 claims description 7
- 208000022831 chronic renal failure syndrome Diseases 0.000 claims description 7
- 230000007882 cirrhosis Effects 0.000 claims description 7
- 208000021921 corneal disease Diseases 0.000 claims description 7
- 230000006378 damage Effects 0.000 claims description 7
- 206010012601 diabetes mellitus Diseases 0.000 claims description 7
- 208000002780 macular degeneration Diseases 0.000 claims description 7
- 244000005700 microbiome Species 0.000 claims description 7
- 208000010125 myocardial infarction Diseases 0.000 claims description 7
- 208000028169 periodontal disease Diseases 0.000 claims description 7
- 208000005069 pulmonary fibrosis Diseases 0.000 claims description 7
- 206010039073 rheumatoid arthritis Diseases 0.000 claims description 7
- 238000012216 screening Methods 0.000 claims description 7
- 208000020431 spinal cord injury Diseases 0.000 claims description 7
- 206010001052 Acute respiratory distress syndrome Diseases 0.000 claims description 6
- 208000013616 Respiratory Distress Syndrome Diseases 0.000 claims description 6
- 201000000028 adult respiratory distress syndrome Diseases 0.000 claims description 6
- 208000008338 non-alcoholic fatty liver disease Diseases 0.000 claims description 6
- 206010053219 non-alcoholic steatohepatitis Diseases 0.000 claims description 6
- 201000000596 systemic lupus erythematosus Diseases 0.000 claims description 6
- 208000011231 Crohn disease Diseases 0.000 claims description 4
- 230000007812 deficiency Effects 0.000 claims 2
- 230000017423 tissue regeneration Effects 0.000 claims 1
- 239000002609 medium Substances 0.000 description 48
- 108020004999 messenger RNA Proteins 0.000 description 47
- 108020004459 Small interfering RNA Proteins 0.000 description 37
- 241000700159 Rattus Species 0.000 description 31
- 210000002966 serum Anatomy 0.000 description 30
- 108090000623 proteins and genes Proteins 0.000 description 25
- 238000012360 testing method Methods 0.000 description 24
- 101000958041 Homo sapiens Musculin Proteins 0.000 description 23
- 101000652133 Homo sapiens STE20-like serine/threonine-protein kinase Proteins 0.000 description 23
- 101000601460 Homo sapiens Serine/threonine-protein kinase Nek4 Proteins 0.000 description 23
- 102100030571 STE20-like serine/threonine-protein kinase Human genes 0.000 description 23
- 102000046299 Transforming Growth Factor beta1 Human genes 0.000 description 23
- 101800002279 Transforming growth factor beta-1 Proteins 0.000 description 23
- 102000018233 Fibroblast Growth Factor Human genes 0.000 description 22
- 108050007372 Fibroblast Growth Factor Proteins 0.000 description 22
- 229940126864 fibroblast growth factor Drugs 0.000 description 22
- 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 description 22
- 108010038512 Platelet-Derived Growth Factor Proteins 0.000 description 21
- 102000010780 Platelet-Derived Growth Factor Human genes 0.000 description 21
- 102000046949 human MSC Human genes 0.000 description 21
- 102000004169 proteins and genes Human genes 0.000 description 21
- 101000934372 Homo sapiens Macrosialin Proteins 0.000 description 19
- 102100025136 Macrosialin Human genes 0.000 description 19
- VBEQCZHXXJYVRD-GACYYNSASA-N uroanthelone Chemical compound C([C@@H](C(=O)N[C@H](C(=O)N[C@@H](CS)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CS)C(=O)N[C@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)NCC(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N[C@@H](CO)C(=O)NCC(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CS)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O)C(C)C)[C@@H](C)O)NC(=O)[C@H](CO)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CO)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@@H](NC(=O)[C@H](CC=1NC=NC=1)NC(=O)[C@H](CCSC)NC(=O)[C@H](CS)NC(=O)[C@@H](NC(=O)CNC(=O)CNC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CS)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)CNC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CO)NC(=O)[C@H](CO)NC(=O)[C@H]1N(CCC1)C(=O)[C@H](CS)NC(=O)CNC(=O)[C@H]1N(CCC1)C(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CO)NC(=O)[C@@H](N)CC(N)=O)C(C)C)[C@@H](C)CC)C1=CC=C(O)C=C1 VBEQCZHXXJYVRD-GACYYNSASA-N 0.000 description 19
- 102000009024 Epidermal Growth Factor Human genes 0.000 description 18
- 101800003838 Epidermal growth factor Proteins 0.000 description 18
- 229940116977 epidermal growth factor Drugs 0.000 description 18
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 17
- LOKCTEFSRHRXRJ-UHFFFAOYSA-I dipotassium trisodium dihydrogen phosphate hydrogen phosphate dichloride Chemical compound P(=O)(O)(O)[O-].[K+].P(=O)(O)([O-])[O-].[Na+].[Na+].[Cl-].[K+].[Cl-].[Na+] LOKCTEFSRHRXRJ-UHFFFAOYSA-I 0.000 description 17
- 206010061218 Inflammation Diseases 0.000 description 15
- 102000004887 Transforming Growth Factor beta Human genes 0.000 description 15
- 108090001012 Transforming Growth Factor beta Proteins 0.000 description 15
- 239000007640 basal medium Substances 0.000 description 15
- 239000002953 phosphate buffered saline Substances 0.000 description 15
- 239000000902 placebo Substances 0.000 description 15
- 229940068196 placebo Drugs 0.000 description 15
- ZRKFYGHZFMAOKI-QMGMOQQFSA-N tgfbeta Chemical compound C([C@H](NC(=O)[C@H](C(C)C)NC(=O)CNC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H]([C@@H](C)O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H]([C@@H](C)O)NC(=O)[C@H](CC(C)C)NC(=O)CNC(=O)[C@H](C)NC(=O)[C@H](CO)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@@H](NC(=O)[C@H](C)NC(=O)[C@H](C)NC(=O)[C@@H](NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](N)CCSC)C(C)C)[C@@H](C)CC)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H](C)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](C)C(=O)N[C@@H](CC(C)C)C(=O)N1[C@@H](CCC1)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(C)C)C(O)=O)C1=CC=C(O)C=C1 ZRKFYGHZFMAOKI-QMGMOQQFSA-N 0.000 description 15
- 102000003745 Hepatocyte Growth Factor Human genes 0.000 description 13
- 108090000100 Hepatocyte Growth Factor Proteins 0.000 description 13
- 230000004054 inflammatory process Effects 0.000 description 13
- 102000017420 CD3 protein, epsilon/gamma/delta subunit Human genes 0.000 description 11
- 102000004877 Insulin Human genes 0.000 description 11
- 108090001061 Insulin Proteins 0.000 description 11
- 241001465754 Metazoa Species 0.000 description 11
- 229940125396 insulin Drugs 0.000 description 11
- 108010071390 Serum Albumin Proteins 0.000 description 10
- 102000007562 Serum Albumin Human genes 0.000 description 10
- UREBDLICKHMUKA-CXSFZGCWSA-N dexamethasone Chemical compound C1CC2=CC(=O)C=C[C@]2(C)[C@]2(F)[C@@H]1[C@@H]1C[C@@H](C)[C@@](C(=O)CO)(O)[C@@]1(C)C[C@@H]2O UREBDLICKHMUKA-CXSFZGCWSA-N 0.000 description 10
- 229960003957 dexamethasone Drugs 0.000 description 10
- 230000002401 inhibitory effect Effects 0.000 description 10
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 9
- 239000012091 fetal bovine serum Substances 0.000 description 9
- 238000012744 immunostaining Methods 0.000 description 9
- 239000013642 negative control Substances 0.000 description 9
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Natural products OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 description 8
- 102000004889 Interleukin-6 Human genes 0.000 description 8
- 108090001005 Interleukin-6 Proteins 0.000 description 8
- 210000001185 bone marrow Anatomy 0.000 description 8
- 239000012888 bovine serum Substances 0.000 description 8
- 230000002829 reductive effect Effects 0.000 description 8
- 102000004625 Aspartate Aminotransferases Human genes 0.000 description 7
- 108010003415 Aspartate Aminotransferases Proteins 0.000 description 7
- 101000856500 Bacillus subtilis subsp. natto Glutathione hydrolase proenzyme Proteins 0.000 description 7
- 230000037396 body weight Effects 0.000 description 7
- 230000006698 induction Effects 0.000 description 7
- 210000001847 jaw Anatomy 0.000 description 7
- 210000003734 kidney Anatomy 0.000 description 7
- 239000000203 mixture Substances 0.000 description 7
- 210000003462 vein Anatomy 0.000 description 7
- 238000001262 western blot Methods 0.000 description 7
- 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 description 6
- 102100021943 C-C motif chemokine 2 Human genes 0.000 description 6
- 101710155857 C-C motif chemokine 2 Proteins 0.000 description 6
- 102100026810 Cyclin-dependent kinase 7 Human genes 0.000 description 6
- 241000282412 Homo Species 0.000 description 6
- 101000911952 Homo sapiens Cyclin-dependent kinase 7 Proteins 0.000 description 6
- 101000932478 Homo sapiens Receptor-type tyrosine-protein kinase FLT3 Proteins 0.000 description 6
- 101000753280 Mus musculus Angiopoietin-1 receptor Proteins 0.000 description 6
- 108060008682 Tumor Necrosis Factor Proteins 0.000 description 6
- 102000000852 Tumor Necrosis Factor-alpha Human genes 0.000 description 6
- 238000003501 co-culture Methods 0.000 description 6
- 230000003247 decreasing effect Effects 0.000 description 6
- 230000012010 growth Effects 0.000 description 6
- 230000001965 increasing effect Effects 0.000 description 6
- 239000000411 inducer Substances 0.000 description 6
- 230000001939 inductive effect Effects 0.000 description 6
- 210000004185 liver Anatomy 0.000 description 6
- VZGDMQKNWNREIO-UHFFFAOYSA-N tetrachloromethane Chemical compound ClC(Cl)(Cl)Cl VZGDMQKNWNREIO-UHFFFAOYSA-N 0.000 description 6
- 108010067225 Cell Adhesion Molecules Proteins 0.000 description 5
- 102000016289 Cell Adhesion Molecules Human genes 0.000 description 5
- 102000008186 Collagen Human genes 0.000 description 5
- 108010035532 Collagen Proteins 0.000 description 5
- 241000699666 Mus <mouse, genus> Species 0.000 description 5
- 230000002159 abnormal effect Effects 0.000 description 5
- 210000004369 blood Anatomy 0.000 description 5
- 239000008280 blood Substances 0.000 description 5
- 230000008859 change Effects 0.000 description 5
- 229920001436 collagen Polymers 0.000 description 5
- 210000002808 connective tissue Anatomy 0.000 description 5
- XEYBRNLFEZDVAW-ARSRFYASSA-N dinoprostone Chemical compound CCCCC[C@H](O)\C=C\[C@H]1[C@H](O)CC(=O)[C@@H]1C\C=C/CCCC(O)=O XEYBRNLFEZDVAW-ARSRFYASSA-N 0.000 description 5
- 229960002986 dinoprostone Drugs 0.000 description 5
- 238000002474 experimental method Methods 0.000 description 5
- 230000004957 immunoregulator effect Effects 0.000 description 5
- 230000008569 process Effects 0.000 description 5
- 230000000770 proinflammatory effect Effects 0.000 description 5
- XEYBRNLFEZDVAW-UHFFFAOYSA-N prostaglandin E2 Natural products CCCCCC(O)C=CC1C(O)CC(=O)C1CC=CCCCC(O)=O XEYBRNLFEZDVAW-UHFFFAOYSA-N 0.000 description 5
- 238000011084 recovery Methods 0.000 description 5
- 239000004017 serum-free culture medium Substances 0.000 description 5
- 238000012453 sprague-dawley rat model Methods 0.000 description 5
- 210000000130 stem cell Anatomy 0.000 description 5
- PORPENFLTBBHSG-MGBGTMOVSA-N 1,2-dihexadecanoyl-sn-glycerol-3-phosphate Chemical compound CCCCCCCCCCCCCCCC(=O)OC[C@H](COP(O)(O)=O)OC(=O)CCCCCCCCCCCCCCC PORPENFLTBBHSG-MGBGTMOVSA-N 0.000 description 4
- 108010076089 accutase Proteins 0.000 description 4
- 239000000654 additive Substances 0.000 description 4
- AWUCVROLDVIAJX-UHFFFAOYSA-N alpha-glycerophosphate Natural products OCC(O)COP(O)(O)=O AWUCVROLDVIAJX-UHFFFAOYSA-N 0.000 description 4
- 230000003110 anti-inflammatory effect Effects 0.000 description 4
- 239000003963 antioxidant agent Substances 0.000 description 4
- 235000006708 antioxidants Nutrition 0.000 description 4
- YZXBAPSDXZZRGB-DOFZRALJSA-N arachidonic acid Chemical compound CCCCC\C=C/C\C=C/C\C=C/C\C=C/CCCC(O)=O YZXBAPSDXZZRGB-DOFZRALJSA-N 0.000 description 4
- 210000001367 artery Anatomy 0.000 description 4
- 235000010323 ascorbic acid Nutrition 0.000 description 4
- 239000011668 ascorbic acid Substances 0.000 description 4
- 229960005070 ascorbic acid Drugs 0.000 description 4
- 210000004556 brain Anatomy 0.000 description 4
- 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 description 4
- 238000001514 detection method Methods 0.000 description 4
- 239000003102 growth factor Substances 0.000 description 4
- 239000001963 growth medium Substances 0.000 description 4
- IPCSVZSSVZVIGE-UHFFFAOYSA-N hexadecanoic acid Chemical compound CCCCCCCCCCCCCCCC(O)=O IPCSVZSSVZVIGE-UHFFFAOYSA-N 0.000 description 4
- 238000002347 injection Methods 0.000 description 4
- 239000007924 injection Substances 0.000 description 4
- WTJKGGKOPKCXLL-RRHRGVEJSA-N phosphatidylcholine Chemical compound CCCCCCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCCCCCCC=CCCCCCCCC WTJKGGKOPKCXLL-RRHRGVEJSA-N 0.000 description 4
- 210000000278 spinal cord Anatomy 0.000 description 4
- 238000001890 transfection Methods 0.000 description 4
- 210000000626 ureter Anatomy 0.000 description 4
- 230000002238 attenuated effect Effects 0.000 description 3
- 238000004113 cell culture Methods 0.000 description 3
- 238000002659 cell therapy Methods 0.000 description 3
- 230000004069 differentiation Effects 0.000 description 3
- 238000011156 evaluation Methods 0.000 description 3
- 238000001943 fluorescence-activated cell sorting Methods 0.000 description 3
- 230000006870 function Effects 0.000 description 3
- 230000002757 inflammatory effect Effects 0.000 description 3
- 239000003112 inhibitor Substances 0.000 description 3
- 238000002360 preparation method Methods 0.000 description 3
- 238000003757 reverse transcription PCR Methods 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- 239000002904 solvent Substances 0.000 description 3
- 210000002303 tibia Anatomy 0.000 description 3
- 230000007704 transition Effects 0.000 description 3
- 210000000689 upper leg Anatomy 0.000 description 3
- OYHQOLUKZRVURQ-NTGFUMLPSA-N (9Z,12Z)-9,10,12,13-tetratritiooctadeca-9,12-dienoic acid Chemical compound C(CCCCCCC\C(=C(/C\C(=C(/CCCCC)\[3H])\[3H])\[3H])\[3H])(=O)O OYHQOLUKZRVURQ-NTGFUMLPSA-N 0.000 description 2
- WRIDQFICGBMAFQ-UHFFFAOYSA-N (E)-8-Octadecenoic acid Natural products CCCCCCCCCC=CCCCCCCC(O)=O WRIDQFICGBMAFQ-UHFFFAOYSA-N 0.000 description 2
- GVJHHUAWPYXKBD-UHFFFAOYSA-N (±)-α-Tocopherol Chemical compound OC1=C(C)C(C)=C2OC(CCCC(C)CCCC(C)CCCC(C)C)(C)CCC2=C1C GVJHHUAWPYXKBD-UHFFFAOYSA-N 0.000 description 2
- TZCPCKNHXULUIY-RGULYWFUSA-N 1,2-distearoyl-sn-glycero-3-phosphoserine Chemical compound CCCCCCCCCCCCCCCCCC(=O)OC[C@H](COP(O)(=O)OC[C@H](N)C(O)=O)OC(=O)CCCCCCCCCCCCCCCCC TZCPCKNHXULUIY-RGULYWFUSA-N 0.000 description 2
- WRGQSWVCFNIUNZ-GDCKJWNLSA-N 1-oleoyl-sn-glycerol 3-phosphate Chemical compound CCCCCCCC\C=C/CCCCCCCC(=O)OC[C@@H](O)COP(O)(O)=O WRGQSWVCFNIUNZ-GDCKJWNLSA-N 0.000 description 2
- LQJBNNIYVWPHFW-UHFFFAOYSA-N 20:1omega9c fatty acid Natural products CCCCCCCCCCC=CCCCCCCCC(O)=O LQJBNNIYVWPHFW-UHFFFAOYSA-N 0.000 description 2
- QSBYPNXLFMSGKH-UHFFFAOYSA-N 9-Heptadecensaeure Natural products CCCCCCCC=CCCCCCCCC(O)=O QSBYPNXLFMSGKH-UHFFFAOYSA-N 0.000 description 2
- 241000283690 Bos taurus Species 0.000 description 2
- 241000283707 Capra Species 0.000 description 2
- 102000015225 Connective Tissue Growth Factor Human genes 0.000 description 2
- 108010039419 Connective Tissue Growth Factor Proteins 0.000 description 2
- 102000004127 Cytokines Human genes 0.000 description 2
- 108090000695 Cytokines Proteins 0.000 description 2
- 241000196324 Embryophyta Species 0.000 description 2
- 241000283073 Equus caballus Species 0.000 description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- 208000004930 Fatty Liver Diseases 0.000 description 2
- 241000282326 Felis catus Species 0.000 description 2
- 108090000379 Fibroblast growth factor 2 Proteins 0.000 description 2
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 description 2
- JZNWSCPGTDBMEW-UHFFFAOYSA-N Glycerophosphorylethanolamin Natural products NCCOP(O)(=O)OCC(O)CO JZNWSCPGTDBMEW-UHFFFAOYSA-N 0.000 description 2
- ZWZWYGMENQVNFU-UHFFFAOYSA-N Glycerophosphorylserin Natural products OC(=O)C(N)COP(O)(=O)OCC(O)CO ZWZWYGMENQVNFU-UHFFFAOYSA-N 0.000 description 2
- 241000699670 Mus sp. Species 0.000 description 2
- 239000005642 Oleic acid Substances 0.000 description 2
- ZQPPMHVWECSIRJ-UHFFFAOYSA-N Oleic acid Natural products CCCCCCCCC=CCCCCCCCC(O)=O ZQPPMHVWECSIRJ-UHFFFAOYSA-N 0.000 description 2
- 241000283973 Oryctolagus cuniculus Species 0.000 description 2
- 235000021314 Palmitic acid Nutrition 0.000 description 2
- 241001494479 Pecora Species 0.000 description 2
- 241000009328 Perro Species 0.000 description 2
- 235000021355 Stearic acid Nutrition 0.000 description 2
- 241000282898 Sus scrofa Species 0.000 description 2
- 108010073929 Vascular Endothelial Growth Factor A Proteins 0.000 description 2
- 102000005789 Vascular Endothelial Growth Factors Human genes 0.000 description 2
- 108010019530 Vascular Endothelial Growth Factors Proteins 0.000 description 2
- ATBOMIWRCZXYSZ-XZBBILGWSA-N [1-[2,3-dihydroxypropoxy(hydroxy)phosphoryl]oxy-3-hexadecanoyloxypropan-2-yl] (9e,12e)-octadeca-9,12-dienoate Chemical compound CCCCCCCCCCCCCCCC(=O)OCC(COP(O)(=O)OCC(O)CO)OC(=O)CCCCCCC\C=C\C\C=C\CCCCC ATBOMIWRCZXYSZ-XZBBILGWSA-N 0.000 description 2
- 238000009825 accumulation Methods 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- 230000000996 additive effect Effects 0.000 description 2
- DTOSIQBPPRVQHS-PDBXOOCHSA-N alpha-linolenic acid Chemical compound CC\C=C/C\C=C/C\C=C/CCCCCCCC(O)=O DTOSIQBPPRVQHS-PDBXOOCHSA-N 0.000 description 2
- 235000020661 alpha-linolenic acid Nutrition 0.000 description 2
- 230000003078 antioxidant effect Effects 0.000 description 2
- 229940114079 arachidonic acid Drugs 0.000 description 2
- 235000021342 arachidonic acid Nutrition 0.000 description 2
- -1 ascorbic acid compound Chemical class 0.000 description 2
- 210000004204 blood vessel Anatomy 0.000 description 2
- 210000004271 bone marrow stromal cell Anatomy 0.000 description 2
- 238000009395 breeding Methods 0.000 description 2
- 230000001488 breeding effect Effects 0.000 description 2
- 210000000845 cartilage Anatomy 0.000 description 2
- 230000010261 cell growth Effects 0.000 description 2
- 230000004663 cell proliferation Effects 0.000 description 2
- 235000012000 cholesterol Nutrition 0.000 description 2
- 239000002285 corn oil Substances 0.000 description 2
- 235000005687 corn oil Nutrition 0.000 description 2
- 210000004087 cornea Anatomy 0.000 description 2
- 210000001508 eye Anatomy 0.000 description 2
- 235000019197 fats Nutrition 0.000 description 2
- 125000005313 fatty acid group Chemical group 0.000 description 2
- 210000004700 fetal blood Anatomy 0.000 description 2
- 230000036541 health Effects 0.000 description 2
- 238000010253 intravenous injection Methods 0.000 description 2
- QXJSBBXBKPUZAA-UHFFFAOYSA-N isooleic acid Natural products CCCCCCCC=CCCCCCCCCC(O)=O QXJSBBXBKPUZAA-UHFFFAOYSA-N 0.000 description 2
- 229960004488 linolenic acid Drugs 0.000 description 2
- KQQKGWQCNNTQJW-UHFFFAOYSA-N linolenic acid Natural products CC=CCCC=CCC=CCCCCCCCC(O)=O KQQKGWQCNNTQJW-UHFFFAOYSA-N 0.000 description 2
- 150000002632 lipids Chemical class 0.000 description 2
- 210000004072 lung Anatomy 0.000 description 2
- 239000003550 marker Substances 0.000 description 2
- 210000000214 mouth Anatomy 0.000 description 2
- 210000003205 muscle Anatomy 0.000 description 2
- WQEPLUUGTLDZJY-UHFFFAOYSA-N n-Pentadecanoic acid Natural products CCCCCCCCCCCCCCC(O)=O WQEPLUUGTLDZJY-UHFFFAOYSA-N 0.000 description 2
- QIQXTHQIDYTFRH-UHFFFAOYSA-N octadecanoic acid Chemical compound CCCCCCCCCCCCCCCCCC(O)=O QIQXTHQIDYTFRH-UHFFFAOYSA-N 0.000 description 2
- OQCDKBAXFALNLD-UHFFFAOYSA-N octadecanoic acid Natural products CCCCCCCC(C)CCCCCCCCC(O)=O OQCDKBAXFALNLD-UHFFFAOYSA-N 0.000 description 2
- ZQPPMHVWECSIRJ-KTKRTIGZSA-N oleic acid Chemical compound CCCCCCCC\C=C/CCCCCCCC(O)=O ZQPPMHVWECSIRJ-KTKRTIGZSA-N 0.000 description 2
- 229960002969 oleic acid Drugs 0.000 description 2
- 235000021313 oleic acid Nutrition 0.000 description 2
- 239000012188 paraffin wax Substances 0.000 description 2
- 210000002379 periodontal ligament Anatomy 0.000 description 2
- 239000000546 pharmaceutical excipient Substances 0.000 description 2
- 210000003800 pharynx Anatomy 0.000 description 2
- 150000008104 phosphatidylethanolamines Chemical class 0.000 description 2
- 150000003905 phosphatidylinositols Chemical class 0.000 description 2
- 210000002826 placenta Anatomy 0.000 description 2
- 230000035755 proliferation Effects 0.000 description 2
- 230000001737 promoting effect Effects 0.000 description 2
- 230000001172 regenerating effect Effects 0.000 description 2
- 201000002793 renal fibrosis Diseases 0.000 description 2
- 238000005070 sampling Methods 0.000 description 2
- 210000003491 skin Anatomy 0.000 description 2
- 238000010186 staining Methods 0.000 description 2
- 239000008117 stearic acid Substances 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 239000004094 surface-active agent Substances 0.000 description 2
- 210000001258 synovial membrane Anatomy 0.000 description 2
- 239000002562 thickening agent Substances 0.000 description 2
- 210000000515 tooth Anatomy 0.000 description 2
- 238000002054 transplantation Methods 0.000 description 2
- 210000005239 tubule Anatomy 0.000 description 2
- 210000003954 umbilical cord Anatomy 0.000 description 2
- 230000000007 visual effect Effects 0.000 description 2
- NMWKYTGJWUAZPZ-WWHBDHEGSA-N (4S)-4-[[(4R,7S,10S,16S,19S,25S,28S,31R)-31-[[(2S)-2-[[(1R,6R,9S,12S,18S,21S,24S,27S,30S,33S,36S,39S,42R,47R,53S,56S,59S,62S,65S,68S,71S,76S,79S,85S)-47-[[(2S)-2-[[(2S)-4-amino-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-amino-3-methylbutanoyl]amino]-3-methylbutanoyl]amino]-3-hydroxypropanoyl]amino]-3-(1H-imidazol-4-yl)propanoyl]amino]-3-phenylpropanoyl]amino]-4-oxobutanoyl]amino]-3-carboxypropanoyl]amino]-18-(4-aminobutyl)-27,68-bis(3-amino-3-oxopropyl)-36,71,76-tribenzyl-39-(3-carbamimidamidopropyl)-24-(2-carboxyethyl)-21,56-bis(carboxymethyl)-65,85-bis[(1R)-1-hydroxyethyl]-59-(hydroxymethyl)-62,79-bis(1H-imidazol-4-ylmethyl)-9-methyl-33-(2-methylpropyl)-8,11,17,20,23,26,29,32,35,38,41,48,54,57,60,63,66,69,72,74,77,80,83,86-tetracosaoxo-30-propan-2-yl-3,4,44,45-tetrathia-7,10,16,19,22,25,28,31,34,37,40,49,55,58,61,64,67,70,73,75,78,81,84,87-tetracosazatetracyclo[40.31.14.012,16.049,53]heptaoctacontane-6-carbonyl]amino]-3-methylbutanoyl]amino]-7-(3-carbamimidamidopropyl)-25-(hydroxymethyl)-19-[(4-hydroxyphenyl)methyl]-28-(1H-imidazol-4-ylmethyl)-10-methyl-6,9,12,15,18,21,24,27,30-nonaoxo-16-propan-2-yl-1,2-dithia-5,8,11,14,17,20,23,26,29-nonazacyclodotriacontane-4-carbonyl]amino]-5-[[(2S)-1-[[(2S)-1-[[(2S)-3-carboxy-1-[[(2S)-1-[[(2S)-1-[[(1S)-1-carboxyethyl]amino]-4-methyl-1-oxopentan-2-yl]amino]-4-methyl-1-oxopentan-2-yl]amino]-1-oxopropan-2-yl]amino]-1-oxopropan-2-yl]amino]-3-(1H-imidazol-4-yl)-1-oxopropan-2-yl]amino]-5-oxopentanoic acid Chemical compound CC(C)C[C@H](NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](C)NC(=O)[C@H](Cc1c[nH]cn1)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@@H]1CSSC[C@H](NC(=O)[C@@H](NC(=O)[C@@H]2CSSC[C@@H]3NC(=O)[C@H](Cc4ccccc4)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@@H](NC(=O)[C@H](Cc4c[nH]cn4)NC(=O)[C@H](CO)NC(=O)[C@H](CC(O)=O)NC(=O)[C@@H]4CCCN4C(=O)[C@H](CSSC[C@H](NC(=O)[C@@H](NC(=O)CNC(=O)[C@H](Cc4c[nH]cn4)NC(=O)[C@H](Cc4ccccc4)NC3=O)[C@@H](C)O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](Cc3ccccc3)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCCCN)C(=O)N3CCC[C@H]3C(=O)N[C@@H](C)C(=O)N2)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC(N)=O)NC(=O)[C@H](Cc2ccccc2)NC(=O)[C@H](Cc2c[nH]cn2)NC(=O)[C@H](CO)NC(=O)[C@@H](NC(=O)[C@@H](N)C(C)C)C(C)C)[C@@H](C)O)C(C)C)C(=O)N[C@@H](Cc2c[nH]cn2)C(=O)N[C@@H](CO)C(=O)NCC(=O)N[C@@H](Cc2ccc(O)cc2)C(=O)N[C@@H](C(C)C)C(=O)NCC(=O)N[C@@H](C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N1)C(=O)N[C@@H](C)C(O)=O NMWKYTGJWUAZPZ-WWHBDHEGSA-N 0.000 description 1
- 208000007082 Alcoholic Fatty Liver Diseases 0.000 description 1
- 206010002091 Anaesthesia Diseases 0.000 description 1
- 229920001342 Bakelite® Polymers 0.000 description 1
- 108010081589 Becaplermin Proteins 0.000 description 1
- 206010065687 Bone loss Diseases 0.000 description 1
- ZAKOWWREFLAJOT-CEFNRUSXSA-N D-alpha-tocopherylacetate Chemical compound CC(=O)OC1=C(C)C(C)=C2O[C@@](CCC[C@H](C)CCC[C@H](C)CCCC(C)C)(C)CCC2=C1C ZAKOWWREFLAJOT-CEFNRUSXSA-N 0.000 description 1
- ZZZCUOFIHGPKAK-UHFFFAOYSA-N D-erythro-ascorbic acid Natural products OCC1OC(=O)C(O)=C1O ZZZCUOFIHGPKAK-UHFFFAOYSA-N 0.000 description 1
- 239000011626 DL-alpha-tocopherylacetate Substances 0.000 description 1
- 235000001809 DL-alpha-tocopherylacetate Nutrition 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 206010015548 Euthanasia Diseases 0.000 description 1
- 108010037362 Extracellular Matrix Proteins Proteins 0.000 description 1
- 102000010834 Extracellular Matrix Proteins Human genes 0.000 description 1
- 102100024785 Fibroblast growth factor 2 Human genes 0.000 description 1
- 102000003974 Fibroblast growth factor 2 Human genes 0.000 description 1
- 108010067306 Fibronectins Proteins 0.000 description 1
- 102000016359 Fibronectins Human genes 0.000 description 1
- 101710107035 Gamma-glutamyltranspeptidase Proteins 0.000 description 1
- 108010010803 Gelatin Proteins 0.000 description 1
- 101710173228 Glutathione hydrolase proenzyme Proteins 0.000 description 1
- 101500025614 Homo sapiens Transforming growth factor beta-1 Proteins 0.000 description 1
- 108090000723 Insulin-Like Growth Factor I Proteins 0.000 description 1
- 102100037850 Interferon gamma Human genes 0.000 description 1
- 108010074328 Interferon-gamma Proteins 0.000 description 1
- PIWKPBJCKXDKJR-UHFFFAOYSA-N Isoflurane Chemical compound FC(F)OC(Cl)C(F)(F)F PIWKPBJCKXDKJR-UHFFFAOYSA-N 0.000 description 1
- 102000003855 L-lactate dehydrogenase Human genes 0.000 description 1
- 108700023483 L-lactate dehydrogenases Proteins 0.000 description 1
- 239000012097 Lipofectamine 2000 Substances 0.000 description 1
- 102100024193 Mitogen-activated protein kinase 1 Human genes 0.000 description 1
- 206010030113 Oedema Diseases 0.000 description 1
- 206010034665 Peritoneal fibrosis Diseases 0.000 description 1
- RVGRUAULSDPKGF-UHFFFAOYSA-N Poloxamer Chemical compound C1CO1.CC1CO1 RVGRUAULSDPKGF-UHFFFAOYSA-N 0.000 description 1
- 239000012980 RPMI-1640 medium Substances 0.000 description 1
- 102000007056 Recombinant Fusion Proteins Human genes 0.000 description 1
- 108010008281 Recombinant Fusion Proteins Proteins 0.000 description 1
- BUGBHKTXTAQXES-UHFFFAOYSA-N Selenium Chemical compound [Se] BUGBHKTXTAQXES-UHFFFAOYSA-N 0.000 description 1
- 102000013275 Somatomedins Human genes 0.000 description 1
- 210000001744 T-lymphocyte Anatomy 0.000 description 1
- 102000004338 Transferrin Human genes 0.000 description 1
- 108090000901 Transferrin Proteins 0.000 description 1
- 102000006747 Transforming Growth Factor alpha Human genes 0.000 description 1
- 101800004564 Transforming growth factor alpha Proteins 0.000 description 1
- 229930003268 Vitamin C Natural products 0.000 description 1
- 229930003427 Vitamin E Natural products 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 230000001154 acute effect Effects 0.000 description 1
- 210000001789 adipocyte Anatomy 0.000 description 1
- 210000004504 adult stem cell Anatomy 0.000 description 1
- 208000026594 alcoholic fatty liver disease Diseases 0.000 description 1
- 230000037005 anaesthesia Effects 0.000 description 1
- 210000004102 animal cell Anatomy 0.000 description 1
- 230000003510 anti-fibrotic effect Effects 0.000 description 1
- 230000000740 bleeding effect Effects 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 239000006172 buffering agent Substances 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- 230000001914 calming effect Effects 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 210000001612 chondrocyte Anatomy 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 239000003636 conditioned culture medium Substances 0.000 description 1
- 238000011109 contamination Methods 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 239000001177 diphosphate Substances 0.000 description 1
- XPPKVPWEQAFLFU-UHFFFAOYSA-J diphosphate(4-) Chemical compound [O-]P([O-])(=O)OP([O-])([O-])=O XPPKVPWEQAFLFU-UHFFFAOYSA-J 0.000 description 1
- 235000011180 diphosphates Nutrition 0.000 description 1
- 238000004043 dyeing Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 210000002744 extracellular matrix Anatomy 0.000 description 1
- 102000006640 gamma-Glutamyltransferase Human genes 0.000 description 1
- WIGCFUFOHFEKBI-UHFFFAOYSA-N gamma-tocopherol Natural products CC(C)CCCC(C)CCCC(C)CCCC1CCC2C(C)C(O)C(C)C(C)C2O1 WIGCFUFOHFEKBI-UHFFFAOYSA-N 0.000 description 1
- 229920000159 gelatin Polymers 0.000 description 1
- 239000008273 gelatin Substances 0.000 description 1
- 235000019322 gelatine Nutrition 0.000 description 1
- 235000011852 gelatine desserts Nutrition 0.000 description 1
- 230000002068 genetic effect Effects 0.000 description 1
- 230000035876 healing Effects 0.000 description 1
- 208000006454 hepatitis Diseases 0.000 description 1
- 231100000283 hepatitis Toxicity 0.000 description 1
- 210000003494 hepatocyte Anatomy 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 230000002718 inhibitory effect on inflammation Effects 0.000 description 1
- 229960002725 isoflurane Drugs 0.000 description 1
- 210000003292 kidney cell Anatomy 0.000 description 1
- 210000005228 liver tissue Anatomy 0.000 description 1
- 206010025135 lupus erythematosus Diseases 0.000 description 1
- 210000004698 lymphocyte Anatomy 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 210000005155 neural progenitor cell Anatomy 0.000 description 1
- 210000000440 neutrophil Anatomy 0.000 description 1
- 238000010899 nucleation Methods 0.000 description 1
- 210000000963 osteoblast Anatomy 0.000 description 1
- 206010033675 panniculitis Diseases 0.000 description 1
- 230000036961 partial effect Effects 0.000 description 1
- 230000037361 pathway Effects 0.000 description 1
- 108010000685 platelet-derived growth factor AB Proteins 0.000 description 1
- 229920001993 poloxamer 188 Polymers 0.000 description 1
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 description 1
- 229920000053 polysorbate 80 Polymers 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 238000003753 real-time PCR Methods 0.000 description 1
- 230000006798 recombination Effects 0.000 description 1
- 230000029058 respiratory gaseous exchange Effects 0.000 description 1
- 230000011664 signaling Effects 0.000 description 1
- 238000003307 slaughter Methods 0.000 description 1
- 210000004003 subcutaneous fat Anatomy 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 230000001629 suppression Effects 0.000 description 1
- 238000001356 surgical procedure Methods 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 208000011580 syndromic disease Diseases 0.000 description 1
- 230000009885 systemic effect Effects 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
- 229940042585 tocopherol acetate Drugs 0.000 description 1
- 239000012096 transfection reagent Substances 0.000 description 1
- 239000012581 transferrin Substances 0.000 description 1
- 235000019154 vitamin C Nutrition 0.000 description 1
- 239000011718 vitamin C Substances 0.000 description 1
- 235000019165 vitamin E Nutrition 0.000 description 1
- 229940046009 vitamin E Drugs 0.000 description 1
- 239000011709 vitamin E Substances 0.000 description 1
Classifications
-
- 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
- A61K35/14—Blood; Artificial blood
- A61K35/19—Platelets; Megacaryocytes
-
- 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
- A61K35/28—Bone marrow; Haematopoietic stem cells; Mesenchymal stem cells of any origin, e.g. adipose-derived stem cells
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
- A61L27/38—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix containing added animal cells
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/02—Stomatological preparations, e.g. drugs for caries, aphtae, periodontitis
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/04—Drugs for disorders of the alimentary tract or the digestive system for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/16—Drugs for disorders of the alimentary tract or the digestive system for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P11/00—Drugs for disorders of the respiratory system
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P13/00—Drugs for disorders of the urinary system
- A61P13/12—Drugs for disorders of the urinary system of the kidneys
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
- A61P17/02—Drugs for dermatological disorders for treating wounds, ulcers, burns, scars, keloids, or the like
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P19/00—Drugs for skeletal disorders
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P19/00—Drugs for skeletal disorders
- A61P19/02—Drugs for skeletal disorders for joint disorders, e.g. arthritis, arthrosis
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P19/00—Drugs for skeletal disorders
- A61P19/04—Drugs for skeletal disorders for non-specific disorders of the connective tissue
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P25/00—Drugs for disorders of the nervous system
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P25/00—Drugs for disorders of the nervous system
- A61P25/28—Drugs for disorders of the nervous system for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P27/00—Drugs for disorders of the senses
- A61P27/02—Ophthalmic agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
- A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
- A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P37/00—Drugs for immunological or allergic disorders
- A61P37/02—Immunomodulators
- A61P37/06—Immunosuppressants, e.g. drugs for graft rejection
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P9/00—Drugs for disorders of the cardiovascular system
- A61P9/10—Drugs for disorders of the cardiovascular system for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
-
- 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
-
- 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
- C12N5/0663—Bone marrow mesenchymal stem cells (BM-MSC)
-
- 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
- C12N5/0667—Adipose-derived stem cells [ADSC]; Adipose stromal stem cells
-
- 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
- C12N2500/42—Organic phosphate, e.g. beta glycerophosphate
-
- 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
- C12N2500/90—Serum-free medium, which may still contain naturally-sourced 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
- C12N2500/99—Serum-free medium
-
- 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
- C12N2501/10—Growth factors
- C12N2501/11—Epidermal growth factor [EGF]
-
- 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
- C12N2501/10—Growth factors
- C12N2501/113—Acidic fibroblast growth factor (aFGF, FGF-1)
-
- 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
- C12N2501/10—Growth factors
- C12N2501/115—Basic fibroblast growth factor (bFGF, FGF-2)
-
- 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
- C12N2501/10—Growth factors
- C12N2501/135—Platelet-derived growth factor [PDGF]
-
- 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
- C12N2501/999—Small molecules not provided for elsewhere
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Animal Behavior & Ethology (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Medicinal Chemistry (AREA)
- Organic Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Biomedical Technology (AREA)
- Immunology (AREA)
- Cell Biology (AREA)
- Developmental Biology & Embryology (AREA)
- Zoology (AREA)
- Biotechnology (AREA)
- Hematology (AREA)
- Epidemiology (AREA)
- Rheumatology (AREA)
- Wood Science & Technology (AREA)
- Genetics & Genomics (AREA)
- Urology & Nephrology (AREA)
- Virology (AREA)
- Dermatology (AREA)
- Transplantation (AREA)
- Gastroenterology & Hepatology (AREA)
- Physical Education & Sports Medicine (AREA)
- Microbiology (AREA)
- General Engineering & Computer Science (AREA)
- Biochemistry (AREA)
- Diabetes (AREA)
- Neurology (AREA)
- Neurosurgery (AREA)
- Pain & Pain Management (AREA)
- Botany (AREA)
Abstract
Description
本発明の一実施形態にかかる生体組織損傷の修復剤の製造方法(以下、「本実施形態の製造方法」ともいう。)は、無血清培地中で間葉系幹細胞を培養する工程を有する方法である。
本実施形態の製造方法においては、間葉系幹細胞を、FGF、PDGF、EGF、少なくとも1つのリン脂質、及び少なくとも1つの脂肪酸を含有する無血清培地、より好ましくは、デキサメタゾン、インスリンおよび血清アルブミンの少なくとも何れか1つをさらに含有する無血清培地において間葉系幹細胞を培養する(以下、第1工程と称する)。なお、無血清培地は、TGF−β、および/または、HGFを含有していないものであってもよい。
本実施形態の製造方法は、上記第1工程の後に、FGF、PDGF、TGF−β、EGF、少なくとも1つのリン脂質、及び少なくとも1つの脂肪酸を含有する無血清培地、より好ましくは、デキサメタゾン、インスリンおよび血清アルブミンの少なくとも何れか1つをさらに含有する無血清培地において、間葉系幹細胞を培養する第2工程をさらに有していてもよい。なお、無血清培地は、HGFを含有していないものであってもよい。
本実施形態の製造方法は、第1工程および/または第2工程の後に、間葉系幹細胞から、造腫瘍性を有していない間葉系幹細胞をスクリーニングするスクリーニング工程をさらに包含していてもよい。
本発明の一実施形態にかかる生体組織損傷の修復剤(以下「本実施形態の修復剤」という。)は、無血清培地中で培養された間葉系幹細胞を含有していることを特徴としている。
本実施形態の修復剤は、TNF−stimulated gene 6 protein(TSG−6)を有効成分として含有していることを特徴としている。
無血清培地で培養した間葉系幹細胞の移植による腎臓線維化の抑制効果に係る検討を行った。
無血清培地で培養した間葉系幹細胞移植による炎症細胞浸潤の抑制効果を調べた。細胞培養条件、及び手法等は、実施例1に記載した通りに行った。Tリンパ球の指標であるCD3およびマクロファージの指標であるCD68を用いて、尿管を結紮したラットの腎臓における炎症細胞浸潤の評価を行った。
無血清培地で培養した骨髄由来間葉系幹細胞による、マクロファージの活性制御能を評価した。
10%MSCsおよびSF−hMSCsを、Transwell systemを用いてTHP−1と共培養した。共培養72時間後にTHP−1を回収し、Immune−regulatory phenotype(M2)マクロファージのマーカーであるCD163およびCD206の発現をFACS法にて定量した。
無血清培地で培養した脂肪由来間葉系幹細胞による、マクロファージの活性制御能を評価した。
10%hAMSCsおよびSF−hAMSCsを、Transwell systemを用いてTHP−1と共培養した。共培養72時間後にTHP−1を回収し、Immune−regulatory phenotype(M2)マクロファージのマーカーであるCD163およびCD206の発現をFACS法にて定量した。
無血清培地で培養した骨髄由来間葉系幹細胞の炎症抑制効果を評価した。線維化の指標であるHGF、腎障害の指標であるPGE2、炎症の指標であるIL−6、および炎症効果抑制作用の指標であるTSG−6を用いて、細胞の炎症抑制効果を確認した。
Sprague Dawleyラット(6〜7週齢)の大腿骨または脛骨の骨髄から採取したMSCsを、無血清培地(STK1またはSTK2)にて単離し、培養して増殖させ、無血清培地培養MSCs(以降SF−MSCsとも称する)を得た。該MCSsに、Negative Control siRNA、またはTSG−6 siRNA(各20nM)をトランスフェクションした。トランスフェクション後の細胞を、作製4日後のラット一側性尿細管結紮モデルの尾静脈へ投与した。ここで、上記Negative Control siRNAをトランスフェクションしたSF−MSCsを投与した群、TSG−6 siRNAをトランスフェクションしたSF−MSCsを投与した群、またはPBSのみを投与した群、尿細管の結紮を行わなかったコントロールの群を各5群ずつ用意した。一側性尿細管結紮術から10日後に各群のラットを屠殺し、腎臓を摘出し、TSG−6の抗炎症効果について確認した。
肝臓の線維化が進むにつれて、肝組織中にはコラーゲンを主成分とする細胞外マトリックスが過剰に沈着し、その終末像として肝臓の繊維化(例えば、肝硬変、ウイルス性肝炎、アルコール性肝炎、非アルコール性脂肪肝炎)に至る。本実施例においては、無血清培地で培養した間葉系幹細胞の移植による肝臓の線維化(例えば、肝硬変、ウイルス性肝炎、アルコール性肝炎、非アルコール性脂肪肝炎)の抑制効果に係る検討を行った。
本試験において、Sprague Dawleyラット(7週齢、オス)の大腿骨または脛骨から、実施例1の記載と同様の方法で、間葉系幹細胞を採取し、当該間葉系幹細胞を培養および継代した。
肝臓の線維化のモデルとして、一般的な、四塩化炭素を用いたモデルラットを作製した。
前記の疾患モデルである「疾患誘導群」に対し、疾患誘導剤の最終投与を行った日(Day31)の翌日(Day32)に、部分採血を行った。
薬剤の投与後、2週間の観察期間を経た後のDay47に、全群に対して、イソフルラン麻酔下で屠殺(放血安楽死)及び採材を行った。採材においては、組織学的評価に供する肝臓の内側葉および外側左葉に加え、生化学的検査に供する血液検体を採取(全採血)した。採取した肝臓の内側葉および外側左葉を、ホルマリンによって固定した後、パラフィンブロックに包埋した。その後、パラフィンブロックを薄切し、シリウスレッド染色標本を作製した。
「疾患誘導群」においては、Day0からDay31までの疾患誘導剤の投与期間に渡って、疾患誘導剤投与直後には、毎回うずくまりが全個体に見られる等、本疾患誘導剤の投与に伴い想定されうる観察所見が見られた。「健常群」においては、そのような観察所見は認められなかった。「細胞投与群」においては、細胞投与後2週間に渡った評価期間において、細胞治療で考慮すべき如何なる異常所見(例えば、拒絶反応等)も認められなかった。
初回投与日(Day0)から、屠殺を行った日(Day47)に至るまでの、各群の体重の推移は、図23に示す通りである。なお、図23、および、後述する図24〜図26において、(a)は「健常群」の試験結果であり、(b)は「偽薬群」の試験結果であり、(c)は「有血清群」の試験結果であり、(d)は「STK群」の試験結果である。
Day47の血液検体から、血清を分離した。以下において、「AST」はアスパラギン酸アミノ基転移酵を、「LDH」は乳酸脱水素酵素を、「γ−GTP」はγグルタミルトランスペプチダーゼを示す。
シリウスレッド染色標本において、各個体の各葉(内側葉および外側左葉の各々)について、中心静脈を中心として、3視野(各個体毎に6視野(=2葉×3視野))の顕微画像を撮像し(対物レンズ10倍)、画像解析装置にて線維化面積を算出した。
本実施例において、「STK群」では、細胞の投与後に、拒絶反応等の細胞治療で考慮すべき異常所見は認められなかった。それ故に、本発明の修復剤は、安全性が高いと考えられる。
Claims (19)
- 無血清培地中で培養された間葉系幹細胞を含有していることを特徴とする、生体組織損傷の修復剤。
- 生体組織の線維化を抑制するためのものであることを特徴とする、請求項1に記載の修復剤。
- 炎症細胞の浸潤を抑制するためのものであることを特徴とする、請求項1に記載の修復剤。
- マクロファージの活性を制御するためのものであることを特徴とする、請求項1に記載の修復剤。
- 上記無血清培地は、FGF、PDGF、EGF、少なくとも1つのリン脂質、及び、少なくとも1つの脂肪酸を含有しているものであることを特徴とする、請求項1〜4の何れか1項に記載の修復剤。
- 上記生体組織損傷は、慢性腎不全、慢性腎臓病、肝硬変、肺線維症、熱傷、間質性肺炎、薬剤性肺炎、放射線肺臓炎、慢性閉塞性肺疾患、急性呼吸促進症候群、軟骨損傷、骨欠損、脊髄損傷、歯周病、多発性硬化症、関節リウマチ、全身性エリテマトーデス、クローン症、糖尿病、動脈硬化、心筋梗塞、脳卒中、アルツハイマー病、黄斑変性症、ウイルス性肝炎、アルコール性肝炎、非アルコール性脂肪肝炎、顎骨再建、口蓋裂、骨置換材、骨欠損、骨系統疾患、ドライアイ、角膜障害、咽頭炎、関節炎、癌、または、線維症に伴う組織損傷であることを特徴とする、請求項1〜5の何れか1項に記載の修復剤。
- 上記間葉系幹細胞は、スクリーニングされた造腫瘍性を有していない間葉系幹細胞であることを特徴とする、請求項1〜6の何れか1項に記載の修復剤。
- 上記間葉系幹細胞は、少なくとも1回継代されたものであることを特徴とする、請求項1〜7の何れか1項に記載の修復剤。
- 上記継代では、哺乳類および微生物由来の成分を含有していない細胞剥離剤を用いて上記間葉系幹細胞を剥離することを特徴とする、請求項8に記載の修復剤。
- 上記間葉系幹細胞は、当該間葉系幹細胞の培養に適した培養容器を用いて培養されたものであることを特徴とする、請求項1〜9の何れか1項に記載の修復剤。
- 無血清培地中で間葉系幹細胞を培養する工程を有することを特徴とする、生体組織損傷の修復剤の製造方法。
- 上記無血清培地は、FGF、PDGF、EGF、少なくとも1つのリン脂質、及び、少なくとも1つの脂肪酸を含有しているものである、請求項11に記載の製造方法。
- 上記培養する工程の後に、造腫瘍性を有していない間葉系幹細胞をスクリーニングする工程を有することを特徴とする、請求項11または12に記載の製造方法。
- 上記培養する工程では、間葉系幹細胞を少なくとも1回継代することを特徴とする、請求項11〜13の何れか1項に記載の製造方法。
- 上記継代では、哺乳類および微生物由来の成分を含有していない細胞剥離剤を用いて上記間葉系幹細胞を剥離することを特徴とする、請求項14に記載の製造方法。
- 上記培養する工程では、上記間葉系幹細胞の培養に適した培養容器を用いて当該間葉系幹細胞を培養することを特徴とする、請求項11〜15の何れか1項に記載の製造方法。
- TSG−6を含有している、生体組織損傷の修復剤であって、
上記生体組織損傷は、慢性腎不全、慢性腎臓病、肝硬変、肺線維症、熱傷、間質性肺炎、薬剤性肺炎、放射線肺臓炎、慢性閉塞性肺疾患、急性呼吸促進症候群、軟骨損傷、骨欠損、脊髄損傷、歯周病、多発性硬化症、関節リウマチ、全身性エリテマトーデス、クローン症、糖尿病、動脈硬化、心筋梗塞、脳卒中、アルツハイマー病、黄斑変性症、ウイルス性肝炎、アルコール性肝炎、非アルコール性脂肪肝炎、顎骨再建、口蓋裂、骨置換材、骨欠損、骨系統疾患、ドライアイ、角膜障害、咽頭炎、関節炎、癌、または、線維症に伴う組織損傷であることを特徴とする修復剤。 - 生体組織の線維化を抑制するためのものであることを特徴とする、請求項17に記載の修復剤。
- 炎症細胞の浸潤を抑制するためのものであることを特徴とする、請求項17に記載の修復剤。
Applications Claiming Priority (5)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP2016256779 | 2016-12-28 | ||
JP2016256779 | 2016-12-28 | ||
JP2017187074 | 2017-09-27 | ||
JP2017187074 | 2017-09-27 | ||
PCT/JP2017/046427 WO2018123968A1 (ja) | 2016-12-28 | 2017-12-25 | 生体組織損傷の修復剤および当該修復剤の製造方法 |
Publications (2)
Publication Number | Publication Date |
---|---|
JPWO2018123968A1 true JPWO2018123968A1 (ja) | 2019-10-31 |
JP7113430B2 JP7113430B2 (ja) | 2022-08-05 |
Family
ID=62711104
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP2018559456A Active JP7113430B2 (ja) | 2016-12-28 | 2017-12-25 | 生体組織損傷の修復剤および当該修復剤の製造方法 |
Country Status (7)
Country | Link |
---|---|
US (2) | US20210128629A1 (ja) |
EP (1) | EP3563857B1 (ja) |
JP (1) | JP7113430B2 (ja) |
KR (1) | KR102654077B1 (ja) |
CN (1) | CN110177560A (ja) |
AU (1) | AU2017388339B2 (ja) |
WO (1) | WO2018123968A1 (ja) |
Families Citing this family (10)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
AU2017388339B2 (en) * | 2016-12-28 | 2024-02-29 | Hiroshima University | Repair agent for living tissue damage and method for producing said repair agent |
EP3591037A4 (en) * | 2017-03-03 | 2020-11-11 | Rohto Pharmaceutical Co., Ltd. | MESENCHYMATIC STEM CELL AND THERAPEUTIC AGENT AGAINST LIVER DISEASES |
EP4035670A4 (en) * | 2019-09-26 | 2023-11-08 | Two Cells Co., Ltd | METHOD FOR PRODUCING A REPAIR AGENT FOR BIOLOGICAL TISSUE DAMAGE AND REPAIR AGENT FOR BIOLOGICAL TISSUE DAMAGE |
CN111297899A (zh) * | 2020-02-21 | 2020-06-19 | 云南雅盛医疗科技有限公司 | 一种脐带间充质干细胞在制备治疗新型冠状肺炎药物中的应用 |
CN111454891A (zh) * | 2020-03-11 | 2020-07-28 | 杭州原生生物科技有限公司 | 一种人胚胎干细胞源性的间充质干细胞的制备及其应用 |
JP2023524150A (ja) * | 2020-05-07 | 2023-06-08 | シンテラ エービー | 呼吸器疾患治療のための幹細胞 |
TWI818286B (zh) * | 2020-07-17 | 2023-10-11 | 南韓商Lg化學股份有限公司 | 包括表現腫瘤壞死因子可誘導基因6的間葉幹細胞之組成物及藥物組成物 |
CN112826833A (zh) * | 2020-07-30 | 2021-05-25 | 中国人民解放军总医院第五医学中心 | 间充质干细胞在制备新冠肺炎引起的肺损伤修复药物中的应用 |
CN115867296A (zh) * | 2020-09-03 | 2023-03-28 | 智再如股份有限公司 | 中枢神经疾病治疗用组合物、中枢神经疾病治疗用组合物的制造方法以及中枢神经疾病治疗用制剂的制造方法 |
WO2022259721A1 (ja) * | 2021-06-10 | 2022-12-15 | 味の素株式会社 | 間葉系幹細胞の製造方法 |
Citations (9)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2007080919A (ja) * | 2005-09-12 | 2007-03-29 | Sanyo Electric Co Ltd | 半導体装置 |
WO2007080919A1 (ja) * | 2006-01-13 | 2007-07-19 | Japan Science And Technology Agency | 動物細胞を無血清培養するための培地用添加剤、キット及びこれらの利用 |
WO2011111787A1 (ja) * | 2010-03-10 | 2011-09-15 | 株式会社ツーセル | 間葉系幹細胞を含む細胞製剤及びその製造方法 |
JP2011525355A (ja) * | 2008-06-18 | 2011-09-22 | ザ テキサス エーアンドエム ユニバーシティ システム | 間充織幹細胞、組成物、及び心臓組織障害の治療方法 |
JP2012157263A (ja) * | 2011-01-31 | 2012-08-23 | Seems Inc | アルツハイマー病治療に向けたヒト脂肪組織由来間葉系幹細胞 |
WO2015016357A1 (ja) * | 2013-08-01 | 2015-02-05 | 株式会社ツーセル | 軟骨損傷治療剤及びその製造方法 |
CN104666347A (zh) * | 2015-02-28 | 2015-06-03 | 广州医科大学附属第一医院 | 脐带间充质干细胞在制备治疗肺间质纤维化的药物制剂中的应用 |
US20160114183A1 (en) * | 2014-10-24 | 2016-04-28 | The Board Of Trustees Of The University Of Illinois | Use of lasers for treating and reversing fibrosis |
JP2016525098A (ja) * | 2013-07-11 | 2016-08-22 | ソウル ナショナル ユニバーシティ ホスピタルSeoul National University Hospital | ヒト胚性幹細胞から由来した間葉系幹細胞を有効成分として含有する肝繊維化または肝硬変予防及び治療用組成物 |
Family Cites Families (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US10046011B2 (en) * | 2008-01-31 | 2018-08-14 | Rutgers, The State University Of New Jersey | Compositions for inducing or suppressing an immune response |
CN102925409B (zh) * | 2012-11-19 | 2014-07-02 | 上海市第六人民医院 | 尿液间充质干细胞的提取及扩增培养方法和应用 |
CN103550117A (zh) * | 2013-10-18 | 2014-02-05 | 上海润生生物技术有限公司 | 可促进皮肤修复与再生的美容护肤品及其制备方法和应用 |
AU2017388339B2 (en) * | 2016-12-28 | 2024-02-29 | Hiroshima University | Repair agent for living tissue damage and method for producing said repair agent |
-
2017
- 2017-12-25 AU AU2017388339A patent/AU2017388339B2/en active Active
- 2017-12-25 CN CN201780081115.XA patent/CN110177560A/zh active Pending
- 2017-12-25 JP JP2018559456A patent/JP7113430B2/ja active Active
- 2017-12-25 US US16/473,987 patent/US20210128629A1/en not_active Abandoned
- 2017-12-25 KR KR1020197021365A patent/KR102654077B1/ko active IP Right Grant
- 2017-12-25 WO PCT/JP2017/046427 patent/WO2018123968A1/ja active Application Filing
- 2017-12-25 EP EP17888752.7A patent/EP3563857B1/en active Active
-
2021
- 2021-10-18 US US17/503,756 patent/US20220031757A1/en active Pending
Patent Citations (9)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2007080919A (ja) * | 2005-09-12 | 2007-03-29 | Sanyo Electric Co Ltd | 半導体装置 |
WO2007080919A1 (ja) * | 2006-01-13 | 2007-07-19 | Japan Science And Technology Agency | 動物細胞を無血清培養するための培地用添加剤、キット及びこれらの利用 |
JP2011525355A (ja) * | 2008-06-18 | 2011-09-22 | ザ テキサス エーアンドエム ユニバーシティ システム | 間充織幹細胞、組成物、及び心臓組織障害の治療方法 |
WO2011111787A1 (ja) * | 2010-03-10 | 2011-09-15 | 株式会社ツーセル | 間葉系幹細胞を含む細胞製剤及びその製造方法 |
JP2012157263A (ja) * | 2011-01-31 | 2012-08-23 | Seems Inc | アルツハイマー病治療に向けたヒト脂肪組織由来間葉系幹細胞 |
JP2016525098A (ja) * | 2013-07-11 | 2016-08-22 | ソウル ナショナル ユニバーシティ ホスピタルSeoul National University Hospital | ヒト胚性幹細胞から由来した間葉系幹細胞を有効成分として含有する肝繊維化または肝硬変予防及び治療用組成物 |
WO2015016357A1 (ja) * | 2013-08-01 | 2015-02-05 | 株式会社ツーセル | 軟骨損傷治療剤及びその製造方法 |
US20160114183A1 (en) * | 2014-10-24 | 2016-04-28 | The Board Of Trustees Of The University Of Illinois | Use of lasers for treating and reversing fibrosis |
CN104666347A (zh) * | 2015-02-28 | 2015-06-03 | 广州医科大学附属第一医院 | 脐带间充质干细胞在制备治疗肺间质纤维化的药物制剂中的应用 |
Non-Patent Citations (2)
Title |
---|
STEM CELL RESEARCH & THERAPY, vol. 6, JPN6018007637, 2015, pages 20 - 14, ISSN: 0004607996 * |
現代医学, vol. 61, no. 2, JPN6018007640, 2013, pages 177 - 184, ISSN: 0004710625 * |
Also Published As
Publication number | Publication date |
---|---|
EP3563857B1 (en) | 2023-04-26 |
WO2018123968A1 (ja) | 2018-07-05 |
EP3563857A1 (en) | 2019-11-06 |
AU2017388339B2 (en) | 2024-02-29 |
EP3563857A4 (en) | 2020-01-15 |
US20220031757A1 (en) | 2022-02-03 |
AU2017388339A1 (en) | 2019-08-01 |
KR20190102015A (ko) | 2019-09-02 |
CN110177560A (zh) | 2019-08-27 |
JP7113430B2 (ja) | 2022-08-05 |
US20210128629A1 (en) | 2021-05-06 |
KR102654077B1 (ko) | 2024-04-04 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
JP7113430B2 (ja) | 生体組織損傷の修復剤および当該修復剤の製造方法 | |
JP6191694B2 (ja) | 軟骨損傷治療剤及びその製造方法 | |
JP6571537B2 (ja) | 療法および予防のための細胞を単離する方法 | |
JP6591434B2 (ja) | 内皮コロニー形成細胞様細胞の生成方法 | |
JP2022527998A (ja) | 高機能な製造されたabcb5+間葉系幹細胞 | |
AU2018376452A1 (en) | Ectodermal mesenchymal stem cells and method for producing same | |
CN105073980A (zh) | 含有适合用于缺血性疾病治疗的细胞的细胞群体的体外增殖方法 | |
CN111417718B (zh) | 由cd31阳性cd45阴性cd200阳性的哺乳动物细胞组成的细胞群及其应用 | |
WO2021060460A1 (ja) | 生体組織損傷の修復剤の製造方法および生体組織損傷の修復剤 | |
WO2020190672A1 (en) | Cardiomyocyte-derived exosomes inducing regeneration of damaged heart tissue | |
JP7444267B2 (ja) | 中枢神経疾患の治療用組成物、中枢神経疾患の治療用組成物の製造方法および中枢神経疾患の治療用製剤の製造方法 | |
WO2024063020A1 (ja) | 椎間板治療剤 | |
CN116437940A (zh) | 骨形成组合物和其用途 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
A621 | Written request for application examination |
Free format text: JAPANESE INTERMEDIATE CODE: A621 Effective date: 20201022 |
|
A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20211005 |
|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20211206 |
|
A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20220222 |
|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20220422 |
|
TRDD | Decision of grant or rejection written | ||
A01 | Written decision to grant a patent or to grant a registration (utility model) |
Free format text: JAPANESE INTERMEDIATE CODE: A01 Effective date: 20220621 |
|
A61 | First payment of annual fees (during grant procedure) |
Free format text: JAPANESE INTERMEDIATE CODE: A61 Effective date: 20220712 |
|
R150 | Certificate of patent or registration of utility model |
Ref document number: 7113430 Country of ref document: JP Free format text: JAPANESE INTERMEDIATE CODE: R150 |