TY - JOUR
T1 - Basic techniques in human mesenchymal stem cell cultures
T2 - Differentiation into osteogenic and adipogenic lineages, genetic perturbations, and phenotypic analyses
AU - Bruedigam, Claudia
AU - van Driel, Marjolein
AU - Koedam, Marijke
AU - van de Peppel, Jeroen
AU - van der Eerden, Bram C.J.
AU - Eijken, Marco
AU - van Leeuwen, Johannes P.T.M.
PY - 2011/6
Y1 - 2011/6
N2 - 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.
AB - 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.
KW - Adipocyte
KW - Alkaline phosphatase
KW - Gene expression analysis
KW - Human mesenchymal stem cell
KW - In vitro differentiation
KW - Lipid vesicle
KW - Mineralization
KW - Osteoblast
UR - http://www.scopus.com/inward/record.url?scp=80053897882&partnerID=8YFLogxK
U2 - 10.1002/9780470151808.sc01h03s17
DO - 10.1002/9780470151808.sc01h03s17
M3 - Article
C2 - 21633940
AN - SCOPUS:80053897882
SN - 1941-7322
VL - 17
SP - 1H.3.1 - 1H.3.20
JO - Current Protocols in Stem Cell Biology
JF - Current Protocols in Stem Cell Biology
IS - 1
ER -