In vitro expansion of human mesenchymal stem cells: choice of serum is a determinant of cell proliferation, differentiation, gene expression, and transcriptome stability.

  • Shahdadfar A
  • Fronsdal K
  • Haug T
 et al. 
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Human bone marrow mesenchymal stem cells (hMSCs) represent an appealing source of adult stem cells for cell therapy and tissue engineering, as they are easily obtained and expanded while maintaining their multilineage differentiation potential. All current protocols for in vitro culture of hMSCs include fetal bovine serum (FBS) as nutritional supplement. FBS is an undesirable additive to cells that are expanded for therapeutic purposes in humans because the use of FBS carries the risk of transmitting viral and prion diseases and proteins that may initiate xenogeneic immune responses. In the present study, we have therefore investigated if autologous serum (AS) or allogeneic human serum (alloHS) could replace FBS for the expansion of hMSCs in vitro. We discovered that the choice of serum affected hMSCs at several different levels. First, hMSCs in AS proliferated markedly faster than hMSCs in FBS, whereas use of alloHS resulted in hMSC growth arrest and death. Second, hMSCs in FBS differentiated more rapidly toward mesenchymal lineages compared with hMSCs in AS. Interestingly, genome-wide microarray analysis identified several transcripts involved in cell cycle and differentiation that were differentially regulated between hMSCs in FBS and AS. Finally, several transcripts, including some involved in cell cycle inhibition, were upregulated in hMSCs in FBS at a late passage, whereas the hMSC transcriptome in AS was remarkably stable. Thus, hMSCs may be expanded rapidly and with stable gene expression in AS in the absence of growth factors, whereas FBS induces a more differentiated and less stable transcriptional profile.

Author-supplied keywords

  • Animals
  • Bone Marrow Cells
  • Cattle
  • Cell Culture Techniques
  • Cell Differentiation
  • Cell Growth Processes
  • Culture Media
  • Gene Expression Profiling
  • Gene Expression Regulation
  • Humans
  • Mesenchymal Stromal Cells
  • Reverse Transcriptase Polymerase Chain Reaction
  • cytology
  • metabolism
  • methods
  • physiology

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  • Aboulghassem Shahdadfar

  • Katrine Fronsdal

  • Terje Haug

  • Finn P Reinholt

  • Jan E Brinchmann

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