TITLE:
The Influence of Culture Medium Type on Cellular Phenotype of Canine Adipose Derived Stem Cells
AUTHORS:
Kristina M. Kiefer, G. Elizabeth Pluhar, Michael G. Conzemius, Timothy D. O’Brien
KEYWORDS:
Canine; Mesenchymal Stem Cells; Adipose-Derived Stromal Cells; Culture Conditions
JOURNAL NAME:
Open Journal of Regenerative Medicine,
Vol.3 No.1,
March
19,
2014
ABSTRACT: Canine adipose derived stem cells (ASCs) hold a great promise for the
therapy of osteoarthritis in veterinary medicine. Current therapy is an
autologous, stromal vascular fraction. Allogeneic ASCs provide many advantages,
including efficient, cost-effective treatments while eliminating a surgical
procedure in a diseased animal. Cultured ASCs can be expanded and
characterized, allowing selection of desirable qualities. Use of allogeneic
ASCs requires selection of a culture medium that provides consistent, desirable
cellular products. The supplements within a medium can greatly influence
cellular phenotypes. We hypothesized that medium type influenced cellular
phenotype, allowing selection of a specified cellular product for clinical applications.
We evaluated ASCs derived from adipose tissue of six dogs, assessing mRNA expression of
proinflammatory: interleukin-1b, cyclooxygenase-2, and anti-inflammatory
mediators: tissue inhibitor metalloproteinase-2 and interleukin -1 receptor
antagonist, via quantitative RT-PCR prior to, and following culture in five
cell culture media: basic cell growth medium (BGM), Keratinocyte N
acetyl-L-cysteine supplemented (KNAC) medium, Multipotent Adult Progenitor Cell
(MAPC) medium, serum free medium (SFM) and xeno-free medium. Major
histocompatability complex I (MHCI), major histocompatability complex II
(MHCII), CD44 and CD90 immunophenotypes were assessed via flow cytometry
analysis. Tri-lineage differentiation (bone, adipose and cartilage tissue) was
utilized to verify multipotency. SFM and xeno-free culture conditions did not
produce cell expansion sufficient to assess phenotype. ASCs prior to culture
had wide variability in all mediator levels, while culturing in the remaining
conditions resulted in more predictable expression levels of inflammatory
mediators, with a decrease in all levels. Cultured ASCs retained expression of
cell surface markers MHCI, CD44 and CD90, while decreasing MHCII expression
levels. KNAC and MAPC medium conditions consistently produced tri-lineage
differentation; BGM, SFM and xeno-free medium did not. Culture condition will
influence phenotype of ASCs, and should be selected according to the intended
therapeutic effect.