Advertisement

Adipogenic Differentiation of Human Mesenchymal Stem Cells

  • Trine Fink
  • Vladimir Zachar
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 698)

Abstract

Mesenchymal stem cells have the capability to differentiate into a number of cell types including adipocytes. The adipocytic phenotype is characterized by intracellular accumulation of lipid droplets as well as transcription of adipocyte-specific genes. This paper details a basic protocol for adipogenic induction of bone marrow and adipose tissue-derived stem cells, as well as protocols for staining lipid accumulation and the transcriptional analysis of PPAR-γ and aP2 by real-time RT-PCR.

Key words

Mesenchymal stem cells Adipogenesis Differentiation Lipid staining PPAR-γ aP2 

References

  1. 1.
    Gregoire, F. M. (2001) Adipocyte differentiation: from fibroblast to endocrine cell. Exp Biol Med (Maywood) 226, 997–1002.Google Scholar
  2. 2.
    MacDougald, O. A., Mandrup S. (2002) Adipogenesis: forces that tip the scales. Trends Endocrinol Metab 13, 5–11.PubMedCrossRefGoogle Scholar
  3. 3.
    Otto, T. C., Lane M. D. (2005) Adipose development: from stem cell to adipocyte. Crit Rev Biochem Mol Biol 40, 229–42.PubMedCrossRefGoogle Scholar
  4. 4.
    Janderova, L., McNeil M., Murrell A. N., Mynatt R. L., Smith S. R. (2003) Human mesenchymal stem cells as an in vitro model for human adipogenesis. Obes Res 11, 65–74.PubMedCrossRefGoogle Scholar
  5. 5.
    Pittenger, M. F., Mackay A. M., Beck S. C., et al. (1999) Multilineage potential of adult human mesenchymal stem cells. Science 284, 143–7.PubMedCrossRefGoogle Scholar
  6. 6.
    Zuk, P. A., Zhu M., Mizuno H., et al. (2001) Multilineage cells from human adipose tissue: implications for cell-based therapies. Tissue Eng 7, 211–28.PubMedCrossRefGoogle Scholar
  7. 7.
    Reger, R. L., Tucker A. H., Wolfe M. R. (2008) Differentiation and characterization of human MSCs. Methods Mol Biol 449, 93–107.PubMedCrossRefGoogle Scholar
  8. 8.
    Bunnell, B. A., Estes B. T., Guilak F., Gimble J. M. (2008) Differentiation of adipose stem cells. Methods Mol Biol 456, 155–71.PubMedCrossRefGoogle Scholar
  9. 9.
    Fink, T., Abildtrup L., Fogd K., et al. (2004) Induction of adipocyte-like phenotype in human mesenchymal stem cells by hypoxia. Stem Cells 22, 1346–55.PubMedCrossRefGoogle Scholar
  10. 10.
    Fink, T., Lund P., Pilgaard L., Rasmussen J. G., Duroux M., Zachar V. (2008) Instability of standard PCR reference genes in adipose-derived stem cells during propagation, differentiation and hypoxic exposure. BMC Mol Biol 9, 98.PubMedCrossRefGoogle Scholar
  11. 11.
    Lund, P., Pilgaard L., Duroux M., Fink T., Zachar V. (2009) Effect of growth media and serum replacements on the proliferation and differentiation of adipose-derived stem cells. Cytotherapy 11, 189–97.PubMedCrossRefGoogle Scholar
  12. 12.
    Vandesompele, J., De Preter K., Pattyn F., et al. (2002) Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genes. Genome Biol 3, RESEARCH0034.PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  1. 1.Laboratory for Stem Cell ResearchAalborg UniversityAalborgDenmark

Personalised recommendations