Effect of serum choice on replicative senescence in mesenchymal stromal cells

Cytotherapy. 2015 Jul;17(7):874-84. doi: 10.1016/j.jcyt.2015.02.012. Epub 2015 Mar 19.

Abstract

Background aims: Multipotent mesenchymal stromal cells (MSCs) are promising candidates for innovative cell therapeutic applications. Before their use, however, they usually need to be expanded in vitro with serum-supplemented media. MSCs can undergo replicative senescence during in vitro expansion, but it is not yet clear how serum supplements influence this process.

Methods: In the present study, we compared how media supplemented with fetal bovine serum (FBS) or calf serum (CS) affected morphology, proliferation, differentiation, senescence and other functional characteristics of human umbilical cord-derived MSCs (UC-MSCs).

Results: UC-MSCs cultured in both FBS- and CS-containing media were able to differentiate along osteogenic and adipogenic lineages but ultimately reached proliferation arrest. However, senescence-associated characteristics, such as β-galactosidase activity, reactive oxygen species levels, proliferation rate and gene expression, demonstrate that UC-MSCs grown with FBS have better proliferation potential and differentiation capacity. In contrast, UC-MSCs grown with CS have a higher proportion of apoptotic cells and senescent characteristics. Possible mechanisms for the observed phenotypes include changes in gene expression (Bax, p16, p21 and p53) and cytokine production (interleukin-6 and interleukin-8).

Conclusions: This study demonstrates that FBS-supplemented media provides a better microenvironment for the expansion of UC-MSCs in vitro than CS-supplemented media. This work provides insight into MSCs generation practices for use in basic research and clinical therapies.

Keywords: calf serum; fetal bovine serum; human umbilical cord–derived mesenchymal stromal cells; replicative senescence.

MeSH terms

  • Animals
  • Apoptosis / drug effects
  • Cattle
  • Cell Culture Techniques / methods*
  • Cell Differentiation / drug effects
  • Cell Proliferation / drug effects
  • Cells, Cultured
  • Cellular Senescence / drug effects*
  • Culture Media / pharmacology*
  • Humans
  • Interleukin-6 / biosynthesis
  • Interleukin-8 / biosynthesis
  • Mesenchymal Stem Cells / cytology
  • Mesenchymal Stem Cells / drug effects*
  • Reactive Oxygen Species / metabolism
  • Serum
  • Umbilical Cord / cytology
  • beta-Galactosidase / metabolism

Substances

  • CXCL8 protein, human
  • Culture Media
  • IL6 protein, human
  • Interleukin-6
  • Interleukin-8
  • Reactive Oxygen Species
  • beta-Galactosidase