Equine marrow-derived mesenchymal stem cells: isolation, differentiation and culture optimization

Document Type: Full paper (Original article)

Authors

1 Department of Stem Cell, Cell Sciences Research Centre, Royan Institute, ACECR, Tehran, Iran

2 Department of Clinical Sciences, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran

3 Department of Clinical Sciences, Faculty of Veterinary Medicine, Ferdowsi University of Mashhad, Mashhad, Iran

Abstract

Most studies regarding the marrow-derived equine mesenchymal stem cells (MSCs) have mainly focused
on the cell transplantation without considering the capacity of differentiation and in vitro requirements of the
cells. These concerns were investigated in the present study. Equine MSCs were isolated from the sternal
marrow aspirates and expanded through two successive subcultures. Passage-2 equine MSC cultures were
then treated with appropriate supplements in order to examine the cell osteogenic, chondrogenic and
adipogenic differentiation potential. Furthermore, the culture of the cells was investigated in terms of the
optimal concentration of fetal bovine serum (FBS) and the initial cell-seeding density. Additionally, a growth
curve was plotted for the cells to study their growth characteristics. According to our findings, equine MSCs
were easily generated specialized bone, cartilage and adipose cell lineages as confirmed by specific staining
and RT-PCR analysis. Moreover, the cells exhibited rapid expansion when being cultivated in the medium
with 15% FBS at 100 cells/cm2. Growth curves indicated that these cells rapidly entered the log phase after a
brief lag (adaptation) period. In summary, marrow-derived equine MSCs possess tripotent differentiation
capacity and rapid growth rate in the appropriate culture conditions.

Keywords