Effect of different serums on culture and growth pattern on equine adipose derived mesenchymal stem cells (hrs-AT MSC)

Abstract

Equine multipotent mesenchymal stem cells (hrs-MSC) can be isolated from various tissues including adipose tissue (AT). We have analyzed the effect of different serum sources on hrs-AT-MSC cultured and assessed proliferation, morphology, viability and immunophenotype and plasticity. The hrs-AT-MSC was cultured separately with growth media containing three different serums: 20% FCS (Gibco), 20% horse serum (Sigma) and 20% allogenic horse serum (Belgium lab) in CO2 incubators. The hrs-AT-MSC growth and proliferation was better in cultural conditions where 20% FCS and 20% horse serum (Sigma) were used. Mesenchymal stem cell count was highest in the condition where horse serum (sigma) was used than both FCS and horse allogenic serum. The viability was more in where allogenic serum (Belgium lab) was used than both FCS (Gibco) and horse serum (Sigma). Like FCS (Gibco), horse serum (Sigma) and allogenic horse serum (Belgium lab) also showed promising /positive effects on equine adipose tissue derived mesenchymal stem cell (hrs-At-MSC) culture and proliferation. Horse serum was found as efficient as fetal calf serum in supporting proliferation and differentiation of equine mesenchymal stem cells in vitro. Further studies are needed to analyze these aspects of MSC in tissue regeneration. Stem cell biology has attracted tremendous interest recently. It is hoped that it will play a major role in the treatment of a number of incurable diseases via transplantation therapy. Several verities of stem cells have been isolated and identified in vivo and in vitro. Very broadly they comprise of two major classes: embryonic and adult mesenchymal stem cells.1 Mesenchymal stem cells (MSCs) because of their self replication, differentiate into various types of mature cells and tissues, and regeneration capabilities are regarded as an excellent source of cells for tissue engineering and for treatment of various incurable diseases and therapeutic uses in gene therapy, drug delivery, and reconstructive surgery.2,3 Recently, induced pluripotent stem cells (iPSC) and embryonic stem cells (ESCs) attracted researchers in organogenesis and cell-mediated therapy experiments, however, teratoma formation, ethical issues, and graft vs host rejection are the major limitations in development and therapeutic application of these cells.4 Due to these limitations, mesenchymal stem cells (MSCs) from adult tissues are now attractive material for and tissue engineering and cell-mediated therapy.5 Isolation of MSC derived from equine species has been reported in a number of different tissues, including bone marrow,6 peripheral blood,7 fat tissue8 and umbilical cord blood.9 Adipose tissue derived equine MSC (hrs-AT-MSC) exhibit the ability to differentiate into different types of cells and tissues in appropriate culture conditions using growth factors and specific hormones into osteoblast, chondroblat and adipocytes and a profound proliferative ability without hampering their own genetic firmness.8 Serum is an integral component for MSC culture in vitro and also it is essential for osteogenic differentiation of MSC along with other factors includes β glycerophosphate, ascorbic acid, and dexamethasone as osteogenic supplements. 10 Serum is an essential component of complete growth media for MSC culture because it provides growth factors, nutrients and extracellular matrix proteins which support MSC cells in vitro.11,12 There is also evidence that serum may act as an antioxidant for cells.13 Despite its zoogenic content, animal serum has been used since the first isolation of MSCs and remains a prime component for their culture and differentiation. Different methods for reducing animal antigens in fetal calf serum (FCS) have been suggested but none alleviate 100% percent risks,14 which leads to current researches for the development of substitute culture conditions, and a move towards the possible use of cheap, readily available as well as potentiate serum from other sources. The purpose of this study was to use three different types of serum for the culture and proliferation of equine adipose tissue derived mesenchymal stem cells (hrs-AT-MSC).