Human mesenchymal stem cells as an in vitro model for human adipogenesis

Obes Res. 2003 Jan;11(1):65-74. doi: 10.1038/oby.2003.11.

Abstract

Objective: To validate the human mesenchymal stem cells (hMSCs) as a new in vitro model for the study of human adipogenesis, to develop the optimal protocol for the differentiation of hMSCs into adipocytes, and to describe effect of mitogen-activated protein kinase on hMSC differentiation into adipocytes.

Research methods and procedures: hMSCs, obtained commercially, were differentiated by exposure to insulin, dexamethasone, indomethacin, and 3-isobutyl-1-methylxanthine three times for 3 days each. Various differentiation conditions were examined to optimize differentiation as measured by Oil Red O staining. The gene expression during adipogenic conversion was assessed by reverse-transcription polymerase chain reaction, real-time reverse-transcription polymerase chain reaction, and Western blotting.

Results: hMSCs differentiated into adipocytes to a different extent depending on the experimental conditions. We have found that differentiation medium based on medium 199 and containing 170 nM insulin, 0.5 mM 3-isobutyl-1-methylxanthine, 0.2 mM indomethacin, 1 microM dexamethasone, and 5% fetal bovine serum was optimal. However, the replacement of fetal bovine serum with rabbit serum (15%) led to further enhancement of differentiation. Inhibition of mitogen-activated protein kinase activation also facilitated adipogenic conversion of hMSCs. The pattern of genes expressed during hMSC differentiation into adipocytes (adipsin, peroxisome proliferator-activated receptor-gamma, CCAAT/enhancer-binding protein-beta, GLUT4, and leptin) was similar to that observed in other in vitro adipocyte models.

Discussion: hMSCs are renewable sources of noncommitted precursors that are able to differentiate into mature adipocytes under the proper hormonal and pharmacological stimuli. Thus, hMSCs represent a new model for the study of human adipogenesis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 1-Methyl-3-isobutylxanthine / pharmacology
  • Adipocytes / cytology*
  • Animals
  • Blood
  • Blotting, Western
  • Cell Differentiation / drug effects
  • Cells, Cultured
  • Clone Cells / cytology
  • Culture Media
  • Cyclin D1 / genetics
  • Dexamethasone / pharmacology
  • Enzyme Activation / drug effects
  • Enzyme Inhibitors / pharmacology
  • Glucocorticoids / pharmacology
  • Glucose / pharmacology
  • Humans
  • Indomethacin / pharmacology
  • Insulin / pharmacology
  • Mesoderm / cytology*
  • Mitogen-Activated Protein Kinase 1 / antagonists & inhibitors
  • Mitogen-Activated Protein Kinase 3
  • Mitogen-Activated Protein Kinases / antagonists & inhibitors
  • Mitogen-Activated Protein Kinases / metabolism
  • Proliferating Cell Nuclear Antigen / genetics
  • RNA, Messenger / analysis
  • Rabbits
  • Reverse Transcriptase Polymerase Chain Reaction
  • Stem Cells / cytology*

Substances

  • Culture Media
  • Enzyme Inhibitors
  • Glucocorticoids
  • Insulin
  • Proliferating Cell Nuclear Antigen
  • RNA, Messenger
  • Cyclin D1
  • Dexamethasone
  • Mitogen-Activated Protein Kinase 1
  • Mitogen-Activated Protein Kinase 3
  • Mitogen-Activated Protein Kinases
  • Glucose
  • 1-Methyl-3-isobutylxanthine
  • Indomethacin