Effects of a defined xeno-free medium on the growth and neurotrophic and angiogenic properties of human adult stem cells

Cytotherapy. 2017 May;19(5):629-639. doi: 10.1016/j.jcyt.2017.02.360.

Abstract

Background: The growth properties and neurotrophic and angiogenic effects of human mesenchymal stromal cells (MSCs) cultured in a defined xeno-free, serum-free medium (MesenCult-XF) were investigated.

Methods: Human MSCs from adipose tissue (ASCs) and bone marrow (BMSCs) were cultured in Minimum Essential Medium-alpha (α-MEM) containing fetal calf serum or in MesenCult-XF. Proliferation was measured over 10 passages and the colony-forming unit (CFU) assay and expression of cluster of differentiation (CD) surface markers were determined. Neurite outgrowth and angiogenic activity of the MSCs were determined.

Results: At early passage, both ASCs and BMSCs showed better proliferation in MesenCult-XF compared with standard α-MEM-containing serum. However, CFUs were significantly lower in MesenCult-XF. ASCs cultured in MesenCult-XF continued to expand at faster rates than cells grown in serum. BMSCs showed morphological changes at late passage in MesenCult-XF and stained positive for senescence β-galactosidase activity. Expression levels of CD73 and CD90 were similar in both cell types under the various culture conditions but CD105 was significantly reduced at passage 10 in MesenCult-XF. In vitro stimulation of the cells enhanced the expression of brain-derived neurotrophic factor (BDNF), vascular endothelial growth factor (VEGF-A) and angiopoietin-1. Stimulated ASCs grown in MesenCult-XF evoked the longest neurite outgrowth in a neuron co-culture model. Stimulated BMSCs grown in MesenCult-XF produced the most extensive network of capillary-like tube structures in an in vitro angiogenesis assay.

Conclusions: ASCs and BMSCs exhibit high levels of neurotrophic and angiogenic activity when grown in the defined serum-free medium indicating their suitability for treatment of various neurological conditions. However, long-term expansion in MesenCult-XF might be restricted to ASCs.

Keywords: adipose; bone marrow; clinical cell culture; mesenchymal stromal cells; neurotrophic factors.

MeSH terms

  • Adipogenesis / drug effects
  • Adipose Tissue / cytology
  • Adult
  • Adult Stem Cells / cytology*
  • Adult Stem Cells / drug effects
  • Cell Proliferation / drug effects
  • Cells, Cultured
  • Culture Media
  • Flow Cytometry
  • Humans
  • Mesenchymal Stem Cells / cytology
  • Neovascularization, Physiologic / drug effects*
  • Nerve Growth Factors / pharmacology*
  • Neurites / drug effects
  • Neurites / metabolism
  • Vascular Endothelial Growth Factor A / metabolism

Substances

  • Culture Media
  • Nerve Growth Factors
  • Vascular Endothelial Growth Factor A