Pooled human platelet lysate versus fetal bovine serum-investigating the proliferation rate, chromosome stability and angiogenic potential of human adipose tissue-derived stem cells intended for clinical use

Cytotherapy. 2013 Sep;15(9):1086-97. doi: 10.1016/j.jcyt.2013.01.217. Epub 2013 Apr 17.

Abstract

Background aims: Because of an increasing focus on the use of adipose-derived stem cells (ASCs) in clinical trials, the culture conditions for these cells are being optimized. We compared the proliferation rates and chromosomal stability of ASCs that had been cultured in Dulbecco's modified Eagle's Medium (DMEM) supplemented with either pooled human platelet lysate (pHPL) or clinical-grade fetal bovine serum (FBS) (DMEM(pHPL) versus DMEM(FBS)).

Methods: ASCs from four healthy donors were cultured in either DMEM(pHPL) or DMEM(FBS), and the population doubling time (PDT) was calculated. ASCs from two of the donors were expanded in DMEM(pHPL) or DMEM(FBS) and cultured for the final week before harvesting with or without the addition of vascular endothelial growth factor. We assessed the chromosomal stability (through the use of array comparative genomic hybridization), the expression of ASC and endothelial surface markers and the differentiation and angiogenic potential of these cells.

Results: The ASCs that were cultured in pHPL exhibited a significantly shorter PDT of 29.6 h (95% confidence interval, 22.3-41.9 h) compared with those cultured in FBS, for which the PDT was 123.9 h (95% confidence interval, 95.6-176.2 h). Comparative genomic hybridization analyses revealed no chromosomal aberrations. Cell differentiation, capillary structure formation and cell-surface marker expression were generally unaffected by the type of medium supplement that was used or by the addition of vascular endothelial growth factor.

Conclusions: We observed that the use of pHPL as a growth supplement for ASCs facilitated a significantly higher proliferation rate compared with FBS without compromising genomic stability or differentiation capacity.

Keywords: adipose tissue–derived stem cells; chromosomal stability; fetal bovine serum; mesenchymal stromal cells; pooled human platelet lysate.

MeSH terms

  • Adipose Tissue / metabolism
  • Adipose Tissue / physiology*
  • Animals
  • Blood Platelets / metabolism*
  • Cattle
  • Cell Culture Techniques / methods
  • Cell Differentiation / physiology
  • Cell Proliferation
  • Cells, Cultured
  • Chromosomal Instability / genetics*
  • Female
  • Humans
  • Neovascularization, Physiologic / physiology*
  • Serum / metabolism*
  • Stem Cells / metabolism
  • Stem Cells / physiology*
  • Vascular Endothelial Growth Factor A / metabolism

Substances

  • VEGFA protein, human
  • Vascular Endothelial Growth Factor A