Equine mesenchymal stem cells from bone marrow, adipose tissue and umbilical cord: immunophenotypic characterization and differentiation potential

Abstract

IntroductionStudies with mesenchymal stem cells (MSCs) are increasing due to their immunomodulatory, anti-inflammatory and tissue regenerative properties. However, there is still no agreement about the best source of equine MSCs for a bank for allogeneic therapy. The aim of this study was to evaluate the cell culture and immunophenotypic characteristics and differentiation potential of equine MSCs from bone marrow (BM-MSCs), adipose tissue (AT-MSCs) and umbilical cord (UC-MSCs) under identical in vitro conditions, to compare these sources for research or an allogeneic therapy cell bank.MethodsThe BM-MSCs, AT-MSCs and UC-MSCs were cultured and evaluated in vitro for their osteogenic, adipogenic and chondrogenic differentiation potential. Additionally, MSCs were assessed for CD105, CD44, CD34, CD90 and MHC-II markers by flow cytometry, and MHC-II was also assessed by immunocytochemistry. To interpret the flow cytometry results, statistical analysis was performed using ANOVA.ResultsThe harvesting and culturing procedures of BM-MSCs, AT-MSCs and UC-MSCs were feasible, with an average cell growth until the third passage of 25 days for BM-MSCs, 15 days for AT-MSCs and 26 days for UC-MSCs. MSCs from all sources were able to differentiate into osteogenic (after 10 days for BM-MSCs and AT-MSCs and 15 days for UC-MSCs), adipogenic (after 8 days for BM-MSCs and AT-MSCs and 15 days for UC-MSCs) and chondrogenic (after 21 days for BM-MSCs, AT-MSCs and UC-MSCs) lineages. MSCs showed high expression of CD105, CD44 and CD90 and low or negative expression of CD34 and MHC-II. The MHC-II was not detected by immunocytochemistry techniques in any of the MSCs studied.ConclusionsThe BM, AT and UC are feasible sources for harvesting equine MSCs, and their immunophenotypic and multipotency characteristics attained minimal criteria for defining MSCs. Due to the low expression of MHC-II by MSCs, all of the sources could be used in clinical trials involving allogeneic therapy in horses. However, the BM-MSCs and AT-MSCs showed fastest ‘‘in vitro’’ differentiation and AT-MSCs showed highest cell growth until third passage. These findings suggest that BM and AT may be preferable for cell banking purposes.

Adipose tissue as mesenchymal stem cells source in equine tendinitis treatment

“…However, recent studies have shown that the differentiation potential of bone marrow progenitor cells and of the adipose tissue is similar (BARBERINI et al, 2014), and that the initially observed differences would due to lack of adjustments in the differentiation inducer medium necessary to the AdMSCs (STRIOGA et al, 2012). It is also worth highlighting that there are no studies comparing the therapeutic effects of MSCs from different sources (adipose tissue x bone marrow) used in equine tendinitis therapy; thus, it is necessary to conduct further investigations about it, since there are signs that the adequate source selection may broaden the effects of the cellular therapy (GARVICAN et al, 2014).…”

“…Since then, many isolation and featuring studies have been conducted (CARVALHO et al, 2009(CARVALHO et al, , 2013bMAMBELLI et al, 2009;BARBERINI et al, 2014). A growing number of studies assessing the impact of stem cells from adipose tissue on large animals have been conducted.…”

Isolation and Characterization of Adipose derived Mesenchymal Stem cells (ADMSCs) in Madras Red Sheep (Ovis aries).

“…The expression of CD markers gets changed during the subsequent cell passages in several species and also due to laboratory procedures, biological differences between the donors because of species, age, gender and site of collection [5,6].…”

Immunophenotypic characterisation and cytogenetic analysis of mesenchymal stem cells from equine bone marrow and foal umbilical cords during in vitro culture

“…The literature offers conflicting data regarding the risk of spontaneous transformation of mesenchymal stem cells during іn vitro culture (2)(3)(4)16). Therefore, investigations of the biological properties of mesenchymal stem cells from equine bone marrow and foal umbilical cords as well as their cytogenetic analysis are highly relevant and timely tasks (13).…”