title: Basic techniques in human mesenchymal stem cell cultures: differentiation into osteogenic and adipogenic lineages, genetic perturbations, and phenotypic analyses.
abstract: 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.
authors.affiliation: Department of Internal Medicine, Erasmus Medical Center, Rotterdam, The Netherlands.
authors.forename: Claudia
authors.initials: C
authors.lastname: Bruedigam
authors.forename: Marjolein van
authors.initials: Mv
authors.lastname: Driel
authors.forename: Marijke
authors.initials: M
authors.lastname: Koedam
authors.forename: Jeroen van de
authors.initials: Jv
authors.lastname: Peppel
authors.forename: Bram C J
authors.initials: BC
authors.lastname: van der Eerden
authors.forename: Marco
authors.initials: M
authors.lastname: Eijken
authors.forename: Johannes P T M
authors.initials: JP
authors.lastname: van Leeuwen
mesh_terms: D050156:Adipogenesis / Q000235:genetics; D000469:Alkaline Phosphatase / Q000378:metabolism; D015415:Biomarkers / Q000378:metabolism; D002113:Calcification, Physiologic; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation / Q000235:genetics; D019070:Cell Lineage; D002478:Cells, Cultured; D004247:DNA / Q000378:metabolism; D018274:Electroporation; D005786:Gene Expression Regulation; D006801:Humans; D008055:Lipids / Q000737:chemistry; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000201:enzymology / Q000378:metabolism*; D010012:Osteogenesis / Q000235:genetics; D010641:Phenotype; D014162:Transfection
chemical_list: D015415:Biomarkers; D008055:Lipids; D004247:DNA; D000469:Alkaline Phosphatase