CA2354978A1 - Culture medium to improve the purity of myoblast culture - Google Patents
Culture medium to improve the purity of myoblast culture Download PDFInfo
- Publication number
- CA2354978A1 CA2354978A1 CA 2354978 CA2354978A CA2354978A1 CA 2354978 A1 CA2354978 A1 CA 2354978A1 CA 2354978 CA2354978 CA 2354978 CA 2354978 A CA2354978 A CA 2354978A CA 2354978 A1 CA2354978 A1 CA 2354978A1
- Authority
- CA
- Canada
- Prior art keywords
- culture
- culture medium
- myoblasts
- valine
- myoblast
- 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.)
- Abandoned
Links
- 210000003098 myoblast Anatomy 0.000 title claims abstract description 24
- 239000001963 growth medium Substances 0.000 title claims abstract description 15
- 229930182831 D-valine Natural products 0.000 claims abstract description 8
- KZSNJWFQEVHDMF-BYPYZUCNSA-N L-valine Chemical compound CC(C)[C@H](N)C(O)=O KZSNJWFQEVHDMF-BYPYZUCNSA-N 0.000 claims abstract description 8
- 229960004295 valine Drugs 0.000 claims abstract description 4
- 239000000203 mixture Substances 0.000 claims description 9
- 102000003974 Fibroblast growth factor 2 Human genes 0.000 claims description 6
- 108090000379 Fibroblast growth factor 2 Proteins 0.000 claims description 6
- 229940024606 amino acid Drugs 0.000 claims description 3
- 150000001413 amino acids Chemical class 0.000 claims description 3
- 125000003625 D-valyl group Chemical group N[C@@H](C(=O)*)C(C)C 0.000 claims 1
- 210000002950 fibroblast Anatomy 0.000 abstract description 8
- KZSNJWFQEVHDMF-SCSAIBSYSA-N D-valine Chemical compound CC(C)[C@@H](N)C(O)=O KZSNJWFQEVHDMF-SCSAIBSYSA-N 0.000 abstract description 7
- 230000012010 growth Effects 0.000 abstract description 2
- 102000018368 Triggering Receptor Expressed on Myeloid Cells-1 Human genes 0.000 description 9
- 108010066451 Triggering Receptor Expressed on Myeloid Cells-1 Proteins 0.000 description 9
- 210000004027 cell Anatomy 0.000 description 8
- 238000002054 transplantation Methods 0.000 description 6
- 101001053946 Homo sapiens Dystrophin Proteins 0.000 description 4
- 239000002609 medium Substances 0.000 description 4
- 210000001087 myotubule Anatomy 0.000 description 4
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 3
- 208000021642 Muscular disease Diseases 0.000 description 3
- 201000009623 Myopathy Diseases 0.000 description 3
- 210000003205 muscle Anatomy 0.000 description 3
- 150000003839 salts Chemical class 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
- 208000031229 Cardiomyopathies Diseases 0.000 description 2
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 2
- XUJNEKJLAYXESH-REOHCLBHSA-N L-Cysteine Chemical compound SC[C@H](N)C(O)=O XUJNEKJLAYXESH-REOHCLBHSA-N 0.000 description 2
- DCXYFEDJOCDNAF-REOHCLBHSA-N L-asparagine Chemical compound OC(=O)[C@@H](N)CC(N)=O DCXYFEDJOCDNAF-REOHCLBHSA-N 0.000 description 2
- WHUUTDBJXJRKMK-VKHMYHEASA-N L-glutamic acid Chemical compound OC(=O)[C@@H](N)CCC(O)=O WHUUTDBJXJRKMK-VKHMYHEASA-N 0.000 description 2
- ROHFNLRQFUQHCH-YFKPBYRVSA-N L-leucine Chemical compound CC(C)C[C@H](N)C(O)=O ROHFNLRQFUQHCH-YFKPBYRVSA-N 0.000 description 2
- KDXKERNSBIXSRK-YFKPBYRVSA-N L-lysine Chemical compound NCCCC[C@H](N)C(O)=O KDXKERNSBIXSRK-YFKPBYRVSA-N 0.000 description 2
- AYFVYJQAPQTCCC-GBXIJSLDSA-N L-threonine Chemical compound C[C@@H](O)[C@H](N)C(O)=O AYFVYJQAPQTCCC-GBXIJSLDSA-N 0.000 description 2
- QIVBCDIJIAJPQS-VIFPVBQESA-N L-tryptophane Chemical compound C1=CC=C2C(C[C@H](N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-VIFPVBQESA-N 0.000 description 2
- OUYCCCASQSFEME-QMMMGPOBSA-N L-tyrosine Chemical compound OC(=O)[C@@H](N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-QMMMGPOBSA-N 0.000 description 2
- AUNGANRZJHBGPY-SCRDCRAPSA-N Riboflavin Chemical compound OC[C@@H](O)[C@@H](O)[C@@H](O)CN1C=2C=C(C)C(C)=CC=2N=C2C1=NC(=O)NC2=O AUNGANRZJHBGPY-SCRDCRAPSA-N 0.000 description 2
- IQFYYKKMVGJFEH-XLPZGREQSA-N Thymidine Chemical compound O=C1NC(=O)C(C)=CN1[C@@H]1O[C@H](CO)[C@@H](O)C1 IQFYYKKMVGJFEH-XLPZGREQSA-N 0.000 description 2
- 230000001413 cellular effect Effects 0.000 description 2
- 239000012091 fetal bovine serum Substances 0.000 description 2
- 102000013361 fetuin Human genes 0.000 description 2
- 108060002885 fetuin Proteins 0.000 description 2
- 238000000338 in vitro Methods 0.000 description 2
- 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 2
- 238000001964 muscle biopsy Methods 0.000 description 2
- GHOKWGTUZJEAQD-UHFFFAOYSA-N pantothenic acid Chemical compound OCC(C)(C)C(O)C(=O)NCCC(O)=O GHOKWGTUZJEAQD-UHFFFAOYSA-N 0.000 description 2
- 230000035755 proliferation Effects 0.000 description 2
- KIDHWZJUCRJVML-UHFFFAOYSA-N putrescine Chemical compound NCCCCN KIDHWZJUCRJVML-UHFFFAOYSA-N 0.000 description 2
- LXNHXLLTXMVWPM-UHFFFAOYSA-N pyridoxine Chemical compound CC1=NC=C(CO)C(CO)=C1O LXNHXLLTXMVWPM-UHFFFAOYSA-N 0.000 description 2
- DAEPDZWVDSPTHF-UHFFFAOYSA-M sodium pyruvate Chemical compound [Na+].CC(=O)C([O-])=O DAEPDZWVDSPTHF-UHFFFAOYSA-M 0.000 description 2
- 238000010186 staining Methods 0.000 description 2
- UCSJYZPVAKXKNQ-HZYVHMACSA-N streptomycin Chemical compound CN[C@H]1[C@H](O)[C@@H](O)[C@H](CO)O[C@H]1O[C@@H]1[C@](C=O)(O)[C@H](C)O[C@H]1O[C@@H]1[C@@H](NC(N)=N)[C@H](O)[C@@H](NC(N)=N)[C@H](O)[C@H]1O UCSJYZPVAKXKNQ-HZYVHMACSA-N 0.000 description 2
- JZRWCGZRTZMZEH-UHFFFAOYSA-N thiamine Chemical compound CC1=C(CCO)SC=[N+]1CC1=CN=C(C)N=C1N JZRWCGZRTZMZEH-UHFFFAOYSA-N 0.000 description 2
- 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 2
- YBJHBAHKTGYVGT-ZKWXMUAHSA-N (+)-Biotin Chemical compound N1C(=O)N[C@@H]2[C@H](CCCCC(=O)O)SC[C@@H]21 YBJHBAHKTGYVGT-ZKWXMUAHSA-N 0.000 description 1
- KZDCMKVLEYCGQX-UDPGNSCCSA-N 2-(diethylamino)ethyl 4-aminobenzoate;(2s,5r,6r)-3,3-dimethyl-7-oxo-6-[(2-phenylacetyl)amino]-4-thia-1-azabicyclo[3.2.0]heptane-2-carboxylic acid;hydrate Chemical compound O.CCN(CC)CCOC(=O)C1=CC=C(N)C=C1.N([C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C(=O)CC1=CC=CC=C1 KZDCMKVLEYCGQX-UDPGNSCCSA-N 0.000 description 1
- 239000001763 2-hydroxyethyl(trimethyl)azanium Substances 0.000 description 1
- QVJPPFAOCXDDPW-UHFFFAOYSA-N 5-[chloro(difluoro)methyl]-1,2-oxazole Chemical compound FC(F)(Cl)C1=CC=NO1 QVJPPFAOCXDDPW-UHFFFAOYSA-N 0.000 description 1
- VVIAGPKUTFNRDU-UHFFFAOYSA-N 6S-folinic acid Natural products C1NC=2NC(N)=NC(=O)C=2N(C=O)C1CNC1=CC=C(C(=O)NC(CCC(O)=O)C(O)=O)C=C1 VVIAGPKUTFNRDU-UHFFFAOYSA-N 0.000 description 1
- 229930024421 Adenine Natural products 0.000 description 1
- GFFGJBXGBJISGV-UHFFFAOYSA-N Adenine Chemical compound NC1=NC=NC2=C1N=CN2 GFFGJBXGBJISGV-UHFFFAOYSA-N 0.000 description 1
- DWRXFEITVBNRMK-UHFFFAOYSA-N Beta-D-1-Arabinofuranosylthymine Natural products O=C1NC(=O)C(C)=CN1C1C(O)C(O)C(CO)O1 DWRXFEITVBNRMK-UHFFFAOYSA-N 0.000 description 1
- 235000019743 Choline chloride Nutrition 0.000 description 1
- AUNGANRZJHBGPY-UHFFFAOYSA-N D-Lyxoflavin Natural products OCC(O)C(O)C(O)CN1C=2C=C(C)C(C)=CC=2N=C2C1=NC(=O)NC2=O AUNGANRZJHBGPY-UHFFFAOYSA-N 0.000 description 1
- CKLJMWTZIZZHCS-UHFFFAOYSA-N D-OH-Asp Natural products OC(=O)C(N)CC(O)=O CKLJMWTZIZZHCS-UHFFFAOYSA-N 0.000 description 1
- QNAYBMKLOCPYGJ-UHFFFAOYSA-N D-alpha-Ala Natural products CC([NH3+])C([O-])=O QNAYBMKLOCPYGJ-UHFFFAOYSA-N 0.000 description 1
- 108010069091 Dystrophin Proteins 0.000 description 1
- 102000001039 Dystrophin Human genes 0.000 description 1
- 102400001368 Epidermal growth factor Human genes 0.000 description 1
- 101800003838 Epidermal growth factor Proteins 0.000 description 1
- 102000004864 Fibroblast growth factor 10 Human genes 0.000 description 1
- 108090001047 Fibroblast growth factor 10 Proteins 0.000 description 1
- 206010016654 Fibrosis Diseases 0.000 description 1
- MPJKWIXIYCLVCU-UHFFFAOYSA-N Folinic acid Natural products NC1=NC2=C(N(C=O)C(CNc3ccc(cc3)C(=O)NC(CCC(=O)O)CC(=O)O)CN2)C(=O)N1 MPJKWIXIYCLVCU-UHFFFAOYSA-N 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- 239000004471 Glycine Substances 0.000 description 1
- 206010019280 Heart failures Diseases 0.000 description 1
- 102000004877 Insulin Human genes 0.000 description 1
- 108090001061 Insulin Proteins 0.000 description 1
- QNAYBMKLOCPYGJ-UWTATZPHSA-N L-Alanine Natural products C[C@@H](N)C(O)=O QNAYBMKLOCPYGJ-UWTATZPHSA-N 0.000 description 1
- CKLJMWTZIZZHCS-UWTATZPHSA-N L-Aspartic acid Natural products OC(=O)[C@H](N)CC(O)=O CKLJMWTZIZZHCS-UWTATZPHSA-N 0.000 description 1
- 235000019766 L-Lysine Nutrition 0.000 description 1
- FFEARJCKVFRZRR-UHFFFAOYSA-N L-Methionine Natural products CSCCC(N)C(O)=O FFEARJCKVFRZRR-UHFFFAOYSA-N 0.000 description 1
- ONIBWKKTOPOVIA-BYPYZUCNSA-N L-Proline Chemical compound OC(=O)[C@@H]1CCCN1 ONIBWKKTOPOVIA-BYPYZUCNSA-N 0.000 description 1
- QNAYBMKLOCPYGJ-REOHCLBHSA-N L-alanine Chemical compound C[C@H](N)C(O)=O QNAYBMKLOCPYGJ-REOHCLBHSA-N 0.000 description 1
- ODKSFYDXXFIFQN-BYPYZUCNSA-N L-arginine Chemical compound OC(=O)[C@@H](N)CCCN=C(N)N ODKSFYDXXFIFQN-BYPYZUCNSA-N 0.000 description 1
- 229930064664 L-arginine Natural products 0.000 description 1
- 235000014852 L-arginine Nutrition 0.000 description 1
- CKLJMWTZIZZHCS-REOHCLBHSA-N L-aspartic acid Chemical compound OC(=O)[C@@H](N)CC(O)=O CKLJMWTZIZZHCS-REOHCLBHSA-N 0.000 description 1
- 235000013878 L-cysteine Nutrition 0.000 description 1
- 239000004201 L-cysteine Substances 0.000 description 1
- ZDXPYRJPNDTMRX-VKHMYHEASA-N L-glutamine Chemical compound OC(=O)[C@@H](N)CCC(N)=O ZDXPYRJPNDTMRX-VKHMYHEASA-N 0.000 description 1
- 229930182816 L-glutamine Natural products 0.000 description 1
- AGPKZVBTJJNPAG-WHFBIAKZSA-N L-isoleucine Chemical compound CC[C@H](C)[C@H](N)C(O)=O AGPKZVBTJJNPAG-WHFBIAKZSA-N 0.000 description 1
- 229930182844 L-isoleucine Natural products 0.000 description 1
- 239000004395 L-leucine Substances 0.000 description 1
- 235000019454 L-leucine Nutrition 0.000 description 1
- FFEARJCKVFRZRR-BYPYZUCNSA-N L-methionine Chemical compound CSCC[C@H](N)C(O)=O FFEARJCKVFRZRR-BYPYZUCNSA-N 0.000 description 1
- 229930195722 L-methionine Natural products 0.000 description 1
- 229930182821 L-proline Natural products 0.000 description 1
- 239000004472 Lysine Substances 0.000 description 1
- 102000018697 Membrane Proteins Human genes 0.000 description 1
- 108010052285 Membrane Proteins Proteins 0.000 description 1
- 101150114843 Mgll gene Proteins 0.000 description 1
- 101001053945 Mus musculus Dystrophin Proteins 0.000 description 1
- DFPAKSUCGFBDDF-UHFFFAOYSA-N Nicotinamide Chemical compound NC(=O)C1=CC=CN=C1 DFPAKSUCGFBDDF-UHFFFAOYSA-N 0.000 description 1
- BELBBZDIHDAJOR-UHFFFAOYSA-N Phenolsulfonephthalein Chemical compound C1=CC(O)=CC=C1C1(C=2C=CC(O)=CC=2)C2=CC=CC=C2S(=O)(=O)O1 BELBBZDIHDAJOR-UHFFFAOYSA-N 0.000 description 1
- 239000005700 Putrescine Substances 0.000 description 1
- 238000011579 SCID mouse model Methods 0.000 description 1
- 239000004473 Threonine Substances 0.000 description 1
- 229930003779 Vitamin B12 Natural products 0.000 description 1
- 229960000643 adenine Drugs 0.000 description 1
- 229960003767 alanine Drugs 0.000 description 1
- 235000001014 amino acid Nutrition 0.000 description 1
- 229960001230 asparagine Drugs 0.000 description 1
- 229960005261 aspartic acid Drugs 0.000 description 1
- IQFYYKKMVGJFEH-UHFFFAOYSA-N beta-L-thymidine Natural products O=C1NC(=O)C(C)=CN1C1OC(CO)C(O)C1 IQFYYKKMVGJFEH-UHFFFAOYSA-N 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 229940098773 bovine serum albumin Drugs 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 229960003178 choline chloride Drugs 0.000 description 1
- SGMZJAMFUVOLNK-UHFFFAOYSA-M choline chloride Chemical compound [Cl-].C[N+](C)(C)CCO SGMZJAMFUVOLNK-UHFFFAOYSA-M 0.000 description 1
- 230000009668 clonal growth Effects 0.000 description 1
- AGVAZMGAQJOSFJ-WZHZPDAFSA-M cobalt(2+);[(2r,3s,4r,5s)-5-(5,6-dimethylbenzimidazol-1-yl)-4-hydroxy-2-(hydroxymethyl)oxolan-3-yl] [(2r)-1-[3-[(1r,2r,3r,4z,7s,9z,12s,13s,14z,17s,18s,19r)-2,13,18-tris(2-amino-2-oxoethyl)-7,12,17-tris(3-amino-3-oxopropyl)-3,5,8,8,13,15,18,19-octamethyl-2 Chemical compound [Co+2].N#[C-].[N-]([C@@H]1[C@H](CC(N)=O)[C@@]2(C)CCC(=O)NC[C@@H](C)OP(O)(=O)O[C@H]3[C@H]([C@H](O[C@@H]3CO)N3C4=CC(C)=C(C)C=C4N=C3)O)\C2=C(C)/C([C@H](C\2(C)C)CCC(N)=O)=N/C/2=C\C([C@H]([C@@]/2(CC(N)=O)C)CCC(N)=O)=N\C\2=C(C)/C2=N[C@]1(C)[C@@](C)(CC(N)=O)[C@@H]2CCC(N)=O AGVAZMGAQJOSFJ-WZHZPDAFSA-M 0.000 description 1
- 229940068840 d-biotin Drugs 0.000 description 1
- 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 1
- 229960003957 dexamethasone Drugs 0.000 description 1
- 210000002889 endothelial cell Anatomy 0.000 description 1
- 229940116977 epidermal growth factor Drugs 0.000 description 1
- 239000000835 fiber Substances 0.000 description 1
- 230000004761 fibrosis Effects 0.000 description 1
- VVIAGPKUTFNRDU-ABLWVSNPSA-N folinic acid Chemical compound C1NC=2NC(N)=NC(=O)C=2N(C=O)C1CNC1=CC=C(C(=O)N[C@@H](CCC(O)=O)C(O)=O)C=C1 VVIAGPKUTFNRDU-ABLWVSNPSA-N 0.000 description 1
- 235000008191 folinic acid Nutrition 0.000 description 1
- 239000011672 folinic acid Substances 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 229960002989 glutamic acid Drugs 0.000 description 1
- 239000003102 growth factor Substances 0.000 description 1
- 229960000367 inositol Drugs 0.000 description 1
- CDAISMWEOUEBRE-GPIVLXJGSA-N inositol Chemical compound O[C@H]1[C@H](O)[C@@H](O)[C@H](O)[C@H](O)[C@@H]1O CDAISMWEOUEBRE-GPIVLXJGSA-N 0.000 description 1
- 229940125396 insulin Drugs 0.000 description 1
- 229960000310 isoleucine Drugs 0.000 description 1
- 229960003136 leucine Drugs 0.000 description 1
- 229960001691 leucovorin Drugs 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 229960004452 methionine Drugs 0.000 description 1
- 238000000034 method Methods 0.000 description 1
- 229960003966 nicotinamide Drugs 0.000 description 1
- 235000005152 nicotinamide Nutrition 0.000 description 1
- 239000011570 nicotinamide Substances 0.000 description 1
- 229960003531 phenolsulfonphthalein Drugs 0.000 description 1
- 229960002429 proline Drugs 0.000 description 1
- 235000008160 pyridoxine Nutrition 0.000 description 1
- 239000011677 pyridoxine Substances 0.000 description 1
- 229960002477 riboflavin Drugs 0.000 description 1
- 235000019192 riboflavin Nutrition 0.000 description 1
- 239000002151 riboflavin Substances 0.000 description 1
- CDAISMWEOUEBRE-UHFFFAOYSA-N scyllo-inosotol Natural products OC1C(O)C(O)C(O)C(O)C1O CDAISMWEOUEBRE-UHFFFAOYSA-N 0.000 description 1
- 229960001153 serine Drugs 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- 210000001057 smooth muscle myoblast Anatomy 0.000 description 1
- 229910000030 sodium bicarbonate Inorganic materials 0.000 description 1
- 229940054269 sodium pyruvate Drugs 0.000 description 1
- RPACBEVZENYWOL-XFULWGLBSA-M sodium;(2r)-2-[6-(4-chlorophenoxy)hexyl]oxirane-2-carboxylate Chemical compound [Na+].C=1C=C(Cl)C=CC=1OCCCCCC[C@]1(C(=O)[O-])CO1 RPACBEVZENYWOL-XFULWGLBSA-M 0.000 description 1
- 229960005322 streptomycin Drugs 0.000 description 1
- 235000019157 thiamine Nutrition 0.000 description 1
- 239000011721 thiamine Substances 0.000 description 1
- 229960002898 threonine Drugs 0.000 description 1
- 229940104230 thymidine Drugs 0.000 description 1
- 235000013619 trace mineral Nutrition 0.000 description 1
- 239000011573 trace mineral Substances 0.000 description 1
- 229960004799 tryptophan Drugs 0.000 description 1
- 229960004441 tyrosine Drugs 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
- 239000011715 vitamin B12 Substances 0.000 description 1
- 235000019163 vitamin B12 Nutrition 0.000 description 1
- 229940011671 vitamin b6 Drugs 0.000 description 1
Classifications
-
- 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
- C12N5/0658—Skeletal muscle cells, e.g. myocytes, myotubes, myoblasts
-
- 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
- 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)
Landscapes
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Life Sciences & Earth Sciences (AREA)
- Biomedical Technology (AREA)
- Genetics & Genomics (AREA)
- Zoology (AREA)
- Organic Chemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Rheumatology (AREA)
- Chemical & Material Sciences (AREA)
- Wood Science & Technology (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Cell Biology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The present invention relates to an improved culture medium that allows the preferential growth of myoblasts over fibroblasts.
Specifically, a culture medium disclosed by Ham et al. was modified by substituting a part or the totality of L-valine with D-valine. This improved culture medium allows the selection of primary myoblasts from fibroblasts, since the fibroblasts cannot use D-valine while the myoblasts can.
Specifically, a culture medium disclosed by Ham et al. was modified by substituting a part or the totality of L-valine with D-valine. This improved culture medium allows the selection of primary myoblasts from fibroblasts, since the fibroblasts cannot use D-valine while the myoblasts can.
Description
TITLE OF THE INVENTION
CULTURE MEDIUM TO IMPROVE THE PURITY OF
MYOBLAST CULTURE
FIELD OF THE INVENTION
The present invention relates to a novel culture medium composition to improve the percentage of myoblasts in a primary culture made from a muscle biopsy, which in turn permits more successful treatment of myopathies and cardiopathies.
BACKGROUND OF THE INVENTION
Myoblast transplantation has been investigated for many years as a potential treatment of myopathies, particularly dystrophies.
Myoblast transplantation is also a potential treatment for various cardiomyopathies and in particular heart insufficiency. Methods to proliferate myoblasts from a muscle biopsy have frequently been published. However, over the years, our research laboratory has developed a culture medium formulation, which permits the enrichment of a myoblast population in a primary culture. This is an important discovery since a low percentage of fibroblasts is better to treat both myopathies and cardiomyopathies without producing an important fibrosis.
Picciano et al. in Exp. Cell. Res. 151:134-147 (1984) reported that D-valine cannot be used by fibroblasts. Endothelial cells were maintained in a medium comprising D-valine while contaminating fibroblasts were removed. Nothing is disclosed in this reference on the capacity of myoblasts to use D-valine.
CULTURE MEDIUM TO IMPROVE THE PURITY OF
MYOBLAST CULTURE
FIELD OF THE INVENTION
The present invention relates to a novel culture medium composition to improve the percentage of myoblasts in a primary culture made from a muscle biopsy, which in turn permits more successful treatment of myopathies and cardiopathies.
BACKGROUND OF THE INVENTION
Myoblast transplantation has been investigated for many years as a potential treatment of myopathies, particularly dystrophies.
Myoblast transplantation is also a potential treatment for various cardiomyopathies and in particular heart insufficiency. Methods to proliferate myoblasts from a muscle biopsy have frequently been published. However, over the years, our research laboratory has developed a culture medium formulation, which permits the enrichment of a myoblast population in a primary culture. This is an important discovery since a low percentage of fibroblasts is better to treat both myopathies and cardiomyopathies without producing an important fibrosis.
Picciano et al. in Exp. Cell. Res. 151:134-147 (1984) reported that D-valine cannot be used by fibroblasts. Endothelial cells were maintained in a medium comprising D-valine while contaminating fibroblasts were removed. Nothing is disclosed in this reference on the capacity of myoblasts to use D-valine.
SUMMARY OF THE INVENTION
The present invention thus relates to a culture medium composition that enhances the proliferation of myoblasts rather than the fibroblasts. We have compared our new culture medium (TREM-1) with a culture medium designed by Dr. Ham for myoblast proliferation (In Vitro Cellular and Developmental Biology. 24(8): 833-844, 1988). The similarities and differences in the compositions of these two culture media are summarized in Tables 1 and 2, respectively. The most important difference between the two media is the replacement of L-Valine in the MCDB120 medium by D-Valine in TREM-1 to enrich myoblast cell populations. We have also eliminated Fetuin in the TREM-1 medium. Another important difference is that we have replaced the Epidermal Growth Factor (EGF) in the MCDB120 by the basic Fibroblast Growth Factor (bFGF) because our laboratory previously demonstrated that bFGF improves the success of myoblast transplantation (see US Patent Number 5,833,978).
Table 1: Common composition of MCDB120 and TREM-1 AMINO ACID mglL
L-Alanine 2.67 L-Arginine.HCl 210.67 L-Asparagine. H20 15.01 L-Aspartic Acid 13.31 L-Cysteine. NCI. H20 35.13 L-Glutamic Acid 4.41 L-Glutamine 1461.50 Glycine 2.25 L-H istidine. HCI. H20 41.93 L-Isoleucine 65.58 L-Leucine 131.17 L-Lysine. HCL 181.65 L-Methionine 29.84 L-Phenylalaline 33.04 L-Proline 11.51 L-Serine 31.53 L-Threonine 35.73 L-Tryptophan 4.08 L-Tyrosine 18.12 VITAMINS
d-Biotin 0.00733 Folinic Acid (Ca salt).5H200.602 DL-alpha-Lipoic Acid 0.002063 Niacinamide 6.11 D-Pantothenic Acid 23.82 (hemi-Ca salt) Pyridoxine. HCL 2.056 Riboflavin 0.003764 Thiamin.HCL 3.373 Vitamin B12 0.01355 OTHER ORGANIC
COMPONENTS
Adenine 0.1351 Choline Chloride 13.96 D-Glucose 1000.00 myo-Inositol 18.016 Putrescine.2HCL 0.0001611 Sodium Pyruvate 110.04 Thymidine 0.02422 BULK INORGANIC SALTS
CaC12.2H20 235.23 KCI 298.20 MgS04.7H20 246.38 NaCI 6430.0 Na2HP04.7H20 134.04 TRACE ELEMENTS
CuS04.5H20 0.002496 FeS04.7H20 0.8340 H2Se03 0.00387 MnS04.5H20 0.000241 Na2Si03.9H20 2.842 (NH4)6Mo7O24.4H20 0.00371 NH4V03 0.000585 NiC12.6H20 0.0000713 ZnS04.7H20 0.08625 BUFFERS, INDICATORS
AND MISCELLENOUS
Phenol Red (Na salt) 1.242 NaHC03 1176.0 Table 2: Differences in composition of MCDB120 and TREM-1 Composition of culture medium TREM-1 MCDB120 per mL per mL
D-Valine 0.117 mg L-Valine 0.117 mg Bovine serum albumin 0.5 mg 0.5 mg Dexamethasone 0.39 0.39 pg pg Insulin 5 pg 180 wg Fetuin 0 mg 0.5 mg Basic Fibroblast Growth Factor10 ng 0 ng (bFGF) Epidermial Growth Factor (EGF)0 ng 10 ng Streptomycin 100 ~,g 100 ~,g Penicillin 100 U 100 U
Fetal bovine serum (FBS) 0.15 0.15 mL mL
To demonstrate the usefulness of the TREM-1 culture medium relative to the MCDB120 culture medium, we prepared myoblast cultures from four (4) healthy individuals. The number of cells after the primary culture and after the first and second passage of the culture was determined by counting cell samples from each individual with an hemacytometer. The percentage of myoblasts in the culture was determined at the same time by staining the cells with a mAb specific for NKH-1, a cell surface protein expressed by the myoblasts but not by the fibroblasts. At each passage, five hundred thousand (500 000) cells were transferred to a 75 cm3 culture flask in order to calculate the number of cells produced at each passage. The results are presented in Table 3. There is a significant increase in the percentage of myoblasts in the culture comprising TREM-1 medium.
The present invention thus relates to a culture medium composition that enhances the proliferation of myoblasts rather than the fibroblasts. We have compared our new culture medium (TREM-1) with a culture medium designed by Dr. Ham for myoblast proliferation (In Vitro Cellular and Developmental Biology. 24(8): 833-844, 1988). The similarities and differences in the compositions of these two culture media are summarized in Tables 1 and 2, respectively. The most important difference between the two media is the replacement of L-Valine in the MCDB120 medium by D-Valine in TREM-1 to enrich myoblast cell populations. We have also eliminated Fetuin in the TREM-1 medium. Another important difference is that we have replaced the Epidermal Growth Factor (EGF) in the MCDB120 by the basic Fibroblast Growth Factor (bFGF) because our laboratory previously demonstrated that bFGF improves the success of myoblast transplantation (see US Patent Number 5,833,978).
Table 1: Common composition of MCDB120 and TREM-1 AMINO ACID mglL
L-Alanine 2.67 L-Arginine.HCl 210.67 L-Asparagine. H20 15.01 L-Aspartic Acid 13.31 L-Cysteine. NCI. H20 35.13 L-Glutamic Acid 4.41 L-Glutamine 1461.50 Glycine 2.25 L-H istidine. HCI. H20 41.93 L-Isoleucine 65.58 L-Leucine 131.17 L-Lysine. HCL 181.65 L-Methionine 29.84 L-Phenylalaline 33.04 L-Proline 11.51 L-Serine 31.53 L-Threonine 35.73 L-Tryptophan 4.08 L-Tyrosine 18.12 VITAMINS
d-Biotin 0.00733 Folinic Acid (Ca salt).5H200.602 DL-alpha-Lipoic Acid 0.002063 Niacinamide 6.11 D-Pantothenic Acid 23.82 (hemi-Ca salt) Pyridoxine. HCL 2.056 Riboflavin 0.003764 Thiamin.HCL 3.373 Vitamin B12 0.01355 OTHER ORGANIC
COMPONENTS
Adenine 0.1351 Choline Chloride 13.96 D-Glucose 1000.00 myo-Inositol 18.016 Putrescine.2HCL 0.0001611 Sodium Pyruvate 110.04 Thymidine 0.02422 BULK INORGANIC SALTS
CaC12.2H20 235.23 KCI 298.20 MgS04.7H20 246.38 NaCI 6430.0 Na2HP04.7H20 134.04 TRACE ELEMENTS
CuS04.5H20 0.002496 FeS04.7H20 0.8340 H2Se03 0.00387 MnS04.5H20 0.000241 Na2Si03.9H20 2.842 (NH4)6Mo7O24.4H20 0.00371 NH4V03 0.000585 NiC12.6H20 0.0000713 ZnS04.7H20 0.08625 BUFFERS, INDICATORS
AND MISCELLENOUS
Phenol Red (Na salt) 1.242 NaHC03 1176.0 Table 2: Differences in composition of MCDB120 and TREM-1 Composition of culture medium TREM-1 MCDB120 per mL per mL
D-Valine 0.117 mg L-Valine 0.117 mg Bovine serum albumin 0.5 mg 0.5 mg Dexamethasone 0.39 0.39 pg pg Insulin 5 pg 180 wg Fetuin 0 mg 0.5 mg Basic Fibroblast Growth Factor10 ng 0 ng (bFGF) Epidermial Growth Factor (EGF)0 ng 10 ng Streptomycin 100 ~,g 100 ~,g Penicillin 100 U 100 U
Fetal bovine serum (FBS) 0.15 0.15 mL mL
To demonstrate the usefulness of the TREM-1 culture medium relative to the MCDB120 culture medium, we prepared myoblast cultures from four (4) healthy individuals. The number of cells after the primary culture and after the first and second passage of the culture was determined by counting cell samples from each individual with an hemacytometer. The percentage of myoblasts in the culture was determined at the same time by staining the cells with a mAb specific for NKH-1, a cell surface protein expressed by the myoblasts but not by the fibroblasts. At each passage, five hundred thousand (500 000) cells were transferred to a 75 cm3 culture flask in order to calculate the number of cells produced at each passage. The results are presented in Table 3. There is a significant increase in the percentage of myoblasts in the culture comprising TREM-1 medium.
5 Table 3: Myoblast Production Individual Nb of % Nb of number cells myoblasts cells myoblasts X X
1,000,000 1,000,000 Individual1Primo 0,66 92 1 76 culture Passage 11,22 89 24 76 Passage 181,76 97 408 87 Individual2Primo 0,66 6 culture Passage 2,5 45 4,88 11 Passage 12 27 84,91 20 Individual3Primo 1,1 94 1,2 91 culture Passage 19,4 99 17,28 93 Passage 426,8 98 387,07 96 Individual4Primo 2,2 71 2 50 culture Passage 58,52 66 35,6 46 Passage 971,43 69 178 69 The formation of muscle fibers following the transplantation of myoblasts grown in each of the two media was compared by transplanting three (3) million cells from each culture into the Tibialis anterior muscle of immunodeficient SCID mice. The number of muscle fibers expressing human dystrophin was established a month after the transplantation by staining cryostat muscle cross-sections with the mAb NCIDys3 (Novocastra inc.) which reacts with human dystrophin but not with mouse dystrophin (results in Table 4). Significantly more muscle fibers expressing human dystrophin appeared following the transplantation of myoblasts grown in TREM-1 than in MCDB120.
Table 4: Muscle Fibers Expressing Human Dystrophin Nb of dystrophin positive Individual fibers number Individual 1 150 64 Individual 2 48 39 Individual 3 93 121 Individual 4 339 81 Although the present invention has been described herein above by way of preferred embodiments thereof, it can be modified without departing from the spirit and nature of the subject invention as defined in the appended claims.
1,000,000 1,000,000 Individual1Primo 0,66 92 1 76 culture Passage 11,22 89 24 76 Passage 181,76 97 408 87 Individual2Primo 0,66 6 culture Passage 2,5 45 4,88 11 Passage 12 27 84,91 20 Individual3Primo 1,1 94 1,2 91 culture Passage 19,4 99 17,28 93 Passage 426,8 98 387,07 96 Individual4Primo 2,2 71 2 50 culture Passage 58,52 66 35,6 46 Passage 971,43 69 178 69 The formation of muscle fibers following the transplantation of myoblasts grown in each of the two media was compared by transplanting three (3) million cells from each culture into the Tibialis anterior muscle of immunodeficient SCID mice. The number of muscle fibers expressing human dystrophin was established a month after the transplantation by staining cryostat muscle cross-sections with the mAb NCIDys3 (Novocastra inc.) which reacts with human dystrophin but not with mouse dystrophin (results in Table 4). Significantly more muscle fibers expressing human dystrophin appeared following the transplantation of myoblasts grown in TREM-1 than in MCDB120.
Table 4: Muscle Fibers Expressing Human Dystrophin Nb of dystrophin positive Individual fibers number Individual 1 150 64 Individual 2 48 39 Individual 3 93 121 Individual 4 339 81 Although the present invention has been described herein above by way of preferred embodiments thereof, it can be modified without departing from the spirit and nature of the subject invention as defined in the appended claims.
REFERENCES
Ham, R.G., St. Clair, J.A., Webster, C and Blau, H.M.
Improved media for normal human muscle satellite cells: Serum-free clonal growth and enhanced growth with low serum. In Vitro Cellular and Developmental Biology 24(8): 833-844, 1988.
Ham, R. G., St-Clair, J.A. and Meyer, S.D. Improved media for rapid clonal growth. Myoblast Transfer Therapy, pp 193-199, Edited by R.
Griggs and G. Karpati. Plenum Press, New York 1990.
Picciano, P.T., Johnson, B., Walenga, R.W., Donovan, M., Borman, B.J., Douglas, W.H.J. and Kreutzer. D.L. Effects of D-valine on pulmonary artery endothelial cells morphology and function in cell culture.
Exp..
Cell Research 151, 134-147, 1984.
Ham, R.G., St. Clair, J.A., Webster, C and Blau, H.M.
Improved media for normal human muscle satellite cells: Serum-free clonal growth and enhanced growth with low serum. In Vitro Cellular and Developmental Biology 24(8): 833-844, 1988.
Ham, R. G., St-Clair, J.A. and Meyer, S.D. Improved media for rapid clonal growth. Myoblast Transfer Therapy, pp 193-199, Edited by R.
Griggs and G. Karpati. Plenum Press, New York 1990.
Picciano, P.T., Johnson, B., Walenga, R.W., Donovan, M., Borman, B.J., Douglas, W.H.J. and Kreutzer. D.L. Effects of D-valine on pulmonary artery endothelial cells morphology and function in cell culture.
Exp..
Cell Research 151, 134-147, 1984.
Claims (2)
1. A composition useful as a culture medium for myoblasts which comprises amino acids, wherein at least a portion of the amino acid L-valine is replaced with D-valine.
2. A composition as defined in claim 1, which further comprises basic Fibroblast Growth Factor (bFGF).
Priority Applications (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CA 2354978 CA2354978A1 (en) | 2001-08-02 | 2001-08-02 | Culture medium to improve the purity of myoblast culture |
| AU2002322245A AU2002322245A1 (en) | 2001-08-02 | 2002-08-02 | Culture medium to improve the purity of myoblast cultures and method using same |
| PCT/CA2002/001216 WO2003012076A2 (en) | 2001-08-02 | 2002-08-02 | Culture medium to improve the purity of myoblast cultures and method using same |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CA 2354978 CA2354978A1 (en) | 2001-08-02 | 2001-08-02 | Culture medium to improve the purity of myoblast culture |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CA2354978A1 true CA2354978A1 (en) | 2003-02-02 |
Family
ID=4169713
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA 2354978 Abandoned CA2354978A1 (en) | 2001-08-02 | 2001-08-02 | Culture medium to improve the purity of myoblast culture |
Country Status (3)
| Country | Link |
|---|---|
| AU (1) | AU2002322245A1 (en) |
| CA (1) | CA2354978A1 (en) |
| WO (1) | WO2003012076A2 (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| ES2235640B1 (en) * | 2003-12-17 | 2006-12-16 | Consejo Sup. Investig. Cientificas | MEANS OF CULTURE OF HUMAN PROGENITING CELLS AND USE FOR THE PROLIFERATION OF SUCH CELLS FOR THEIR AUTOMOTIVE TRANSPLANT. |
| EP3406704B1 (en) * | 2016-11-01 | 2021-10-20 | Terumo Kabushiki Kaisha | Modification method for adhesion-state cell culture |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5143842A (en) * | 1988-11-01 | 1992-09-01 | The University Of Colorado Foundation, Inc. | Media for normal human muscle satellite cells |
-
2001
- 2001-08-02 CA CA 2354978 patent/CA2354978A1/en not_active Abandoned
-
2002
- 2002-08-02 AU AU2002322245A patent/AU2002322245A1/en not_active Abandoned
- 2002-08-02 WO PCT/CA2002/001216 patent/WO2003012076A2/en not_active Application Discontinuation
Also Published As
| Publication number | Publication date |
|---|---|
| WO2003012076A3 (en) | 2003-05-30 |
| WO2003012076A2 (en) | 2003-02-13 |
| AU2002322245A1 (en) | 2003-02-17 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US5324656A (en) | Media for normal human muscle satellite cells | |
| JP2577114B2 (en) | Cell culture media | |
| KR101138091B1 (en) | Method for manufacturing basic culture media for mesenchymal stem cell, basic culture media for mesenchymal stem cell and cell theraphy products cultured and differenciated using the same | |
| US6372210B2 (en) | Method for repopulating human bone marrow comprising culturing CD34+ cells in a serum free medium | |
| US7169610B2 (en) | Serum-free media for chondrocytes and methods of use thereof | |
| US7462487B2 (en) | Cell culture media | |
| US6692961B1 (en) | Defined systems for epithelial cell culture and use thereof | |
| US7449334B2 (en) | Medium containing pipecholic acid and gamma amino butyric acid and culture of embryonic stem cells | |
| KR20070058584A (en) | Culturing human embryonic stem cells | |
| US20080124801A1 (en) | Pluripotent cell growth media | |
| TW200927927A (en) | Stem cell medium | |
| AU2007274065A1 (en) | Cell growth medium | |
| JP2000506374A5 (en) | ||
| KR19990071817A (en) | Improved immortalized human skin cell lines and novel serum-free media useful for their preparation | |
| AU756151B2 (en) | Immortalised cell lines derived from normal human skin tissues | |
| CA2504179A1 (en) | Composition for culturing multipotent stem cells and utilization of the same | |
| US20050019310A1 (en) | Method for culturing and expansion of mammalian undifferentiated epidermal kerainocytes exhibiting stem cell characteristics | |
| CA1221651A (en) | Cell growth medium supplement | |
| CA2354978A1 (en) | Culture medium to improve the purity of myoblast culture | |
| IL292933A (en) | Serum-free human pluripotent stem cell culture medium | |
| Agy et al. | Protein-free medium for C-1300 mouse neuroblastoma cells | |
| CN112342187A (en) | Chondrocyte culture medium and preparation method thereof | |
| US5916809A (en) | Medium for culturing normal human epidermal melanocytes | |
| US20240150715A1 (en) | Optimized cell culture medium utilizing iron (III) citrate as an iron delivery method for the in vitro, bioreactor-centric production of manufactured blood | |
| Kumar | Growth of neural cell cultures in a chemically defined, serum-free culture medium |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| FZDE | Dead |