This unit describes basic techniques in human mesenchymal stem cell (hMSC) cultures. It includes protocols for the differentiation of hMSCs into osteogenic and adipogenic lineages, genetic perturbations, and phenotypic analyses. hMSCs can be differentiated with dexamethasone and β-glycerophosphate into mineralizing osteoblasts within 2 to 3 weeks, or with dexamethasone, indomethacin, and 3-isobutyl-1-methylxanthine into lipid vesicle-containing adipocytes within 1 to 2 weeks. Phenotypic changes during those highly dynamic differentiation processes can be detected by biochemical and histological assays and gene expression analyses of differentiation markers. In addition, this unit describes an electroporation method that allows the transient genetic perturbation of hMSCs.

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doi.org/10.1002/9780470151808.sc01h03s17, hdl.handle.net/1765/74905
American Journal of Medical Genetics. Part B: Neuropsychiatric Genetics
Department of Internal Medicine

Bruedigam, C., van Driel, M., Koedam, M., van de Peppel, J., van der Eerden, B., Eijken, M., & van Leeuwen, H. (2011). Basic techniques in human mesenchymal stem cell cultures: Differentiation into osteogenic and adipogenic lineages, genetic perturbations, and phenotypic analyses. American Journal of Medical Genetics. Part B: Neuropsychiatric Genetics, (SUPPL. 17). doi:10.1002/9780470151808.sc01h03s17