Large-scale expansion of pre-isolated bone marrow mesenchymal stromal cells in serum-free conditions

J Tissue Eng Regen Med. 2016 Feb;10(2):108-19. doi: 10.1002/term.1713. Epub 2013 Mar 12.

Abstract

The regenerative potential of mesenchymal stromal or stem cells (MSCs) has generated tremendous interest for treating various degenerative diseases. Regulatory preference is to use a culture medium that is devoid of bovine components for stem cell expansion intended for therapeutic applications. However, a clear choice an alternative to fetal bovine serum (FBS) has not yet emerged. We have screened five different commercially available serum-free media (SFM) for their ability to support the growth and expansion of pre-isolated undifferentiated bone marrow-derived MSCs (BM-MSCs) and compared the results with cells grown in standard FBS-containing medium as control. In addition, based on initial screening results, BD Mosaic™ Mesenchymal Stem Cell Serum-free (BD-SFM) medium was evaluated in large-scale cultures for the performance and culture characteristics of BM-MSCs. Of the five different serum-free media, BD-SFM enhanced BM-MSCs growth and expansion in Cell STACK (CS), but the cell yield per CS-10 was less when compared to the control medium. The characteristics of MSCs were measured in terms of population doubling time (PDT), cell yield and expression of MSC-specific markers. Significant differences were observed between BD-SFM and control medium in terms of population doublings (PDs), cell yield, CFU-F and morphological features, whereas surface phenotype and differentiation potentials were comparable. The BD-SFM-cultured MSCs were also found to retain the differentiation potential, immune-privileged status and immunosuppressive properties inherent to MSCs. Our results suggest that BD-SFM supports large-scale expansion of BM-MSCs for therapeutic use.

Keywords: bone marrow; differentiation; large-scale expansion; mesenchymal stromal cells; phenotypic characterization; serum-free cultures.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biological Assay
  • Bone Marrow Cells / cytology*
  • Cell Differentiation
  • Cell Proliferation
  • Cell Separation / methods*
  • Cell Shape
  • Colony-Forming Units Assay
  • Culture Media, Serum-Free
  • Humans
  • Immunomodulation
  • Immunophenotyping
  • Immunosuppression Therapy
  • Kinetics
  • Mesenchymal Stem Cells / cytology*
  • Serum Albumin, Bovine / metabolism
  • Young Adult

Substances

  • Culture Media, Serum-Free
  • Serum Albumin, Bovine