Differentiation and characterization of human MSCs

Center for Gene Therapy, Tulane University Health Sciences Center, New Orleans, LA, USA.
Methods in Molecular Biology (Impact Factor: 1.29). 02/2008; 449:93-107. DOI: 10.1007/978-1-60327-169-1_7
Source: PubMed

ABSTRACT One of the hallmark characteristics of human MSCs (hMSCs) is their ability to differentiate into adipocytes, chondrocytes and osteocytes in culture. The default fate for hMSCs appears to be bone: if late-passage cultures are left in basic culture medium, the hMSCs will become confluent and produce mineral, an indication of bone formation. However, when grown under certain culture conditions or in media containing specific components, the cells can be driven to become a number of other specific cell types including neural cells, myocytes, and cardiomyocytes. The protocols given here are the basic differentiation procedures for inducing osteogenesis, adipogenesis, and chondrogenesis in cultures of hMSCs. Although there is still no clear consensus on the antigen expression pattern that will define hMSCs, a protocol is also presented for the flow cytometric analysis using a series of antibody panels. The analysis of these surface epitope patterns can aide in the isolation and characterization of hMSCs.

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Available from: Hugh Alan Tucker, Feb 17, 2014
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    • "(CCM) comprising a-Minimum Essential Medium (a-MEM) supplemented with 16.5% (v/v) fetal bovine serum (FBS, Atlanta Biologicals, Inc., Lawrenceville, GA, USA), 2 mM L-glutamine (Gibco/Invitrogen, Carlsbad, CA, USA), 100 U/ml penicillin, and 100 mg/ml streptomycin (Gibco/Invitrogen, Carlsbad, CA, USA). The cells were seeded at a density of 3000 cells/cm 2 and placed in an incubator under 5% CO 2 [29] [30] "
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    • "When the cells reached 70% confluence, cell medium was aspirated and appropriated induction media (set this time point as day 0) were introduced. The induced differentiation treatment methods were used as previously described [12]. Briefly, adipogenesis medium was complete medium supplement with a cocktail of 0.5 lM dexamethasone, 0.5 lM isobutylmethylxanthine , and 50 lM indomethacin. "
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