Abstract
IntroductionMesenchymal stromal cells (MSCs) have been extensively studied for their promising capabilities in regenerative medicine. Although bone marrow is the best-known source for isolating equine MSCs, non-invasive alternative sources such as umbilical cord blood (UCB), umbilical cord matrix (UCM), and peripheral blood (PB) have also been reported.MethodsEquine MSCs from three non-invasive alternative sources were isolated from six individual mares (PB) and their foals (UCB and UCM) at parturition. To minimize inter-horse variability, the samples from the three sources were matched within the same mare and for UCB and UCM even within the same foal from that specific mare. The following parameters were analyzed: (i) success rate of isolation, (ii) proliferation capacity, (iii) tri-lineage differentiation ability, (iv) immunophenotypical protein, and (v) immunomodulatory mRNA profiles. Linear regression models were fit to determine the association between the source of MSCs (UCB, UCM, PB) and (i) the moment of first observation, (ii) the moment of first passage, (iii) cell proliferation data, (iv) the expression of markers related to cell immunogenicity, and (v) the mRNA profile of immunomodulatory factors, except for hepatocyte growth factor (HGF) as no normal distribution could be obtained for the latter variable. To evaluate the association between the source of MSCs and the mRNA expression of HGF, the non-parametric Kruskal-Wallis test was performed instead.ResultsWhile equine MSCs could be isolated from all the UCB and PB samples, isolation from UCM was successful in only two samples because of contamination issues. Proliferation data showed that equine MSCs from all three sources could be easily expanded, although UCB-derived MSCs appeared significantly faster in culture than PB- or UCM-derived MSCs. Equine MSCs from both UCB and PB could be differentiated toward the osteo-, chondro-, and adipogenic lineage, in contrast to UCM-derived MSCs in which only chondro- and adipogenic differentiation could be confirmed. Regardless of the source, equine MSCs expressed the immunomodulatory genes CD40, CD80, HGF, and transforming growth factor-beta (TGFβ). In contrast, no mRNA expression was found for CD86, indoleamine 2,3-dioxygenase (IDO), and tumor necrosis factor-alpha (TNFα).ConclusionsWhereas UCM seems less feasible because of the high contamination risks and low isolation success rates, UCB seems a promising alternative MSC source, especially when considering allogeneic MSC use.
Highlights
Mesenchymal stromal cells (MSCs) have been extensively studied for their promising capabilities in regenerative medicine
Linear regression models were fit to determine the association between the source of MSCs (UCB, umbilical cord matrix (UCM), peripheral blood (PB)) and (i) the moment of first observation, (ii) the moment of first passage, (iii) cell proliferation data, (iv) the expression of markers related to cell immunogenicity, and (v) the mRNA profile of all immunomodulatory factors, except for hepatocyte growth factor (HGF) as no normal distribution could be obtained for the latter variable
Success rate of isolating equine mesenchymal stromal cells umbilical cord blood (UCB), UCM, and PB samples were collected from six mares with a normal parturition, and putative equine MSCs could be isolated from all three sources (Table 3)
Summary
Mesenchymal stromal cells (MSCs) have been extensively studied for their promising capabilities in regenerative medicine. A study by Guest and colleagues [9] found that the injection of allogeneic bone marrow (BM)-derived MSCs did not cause any observable cell-mediated immune response from the host. In a later study by Carrade and colleagues [10], no significant differences could be demonstrated either in the type or in the degree of inflammation when using autologous or related or unrelated allogeneic cells. These findings indicate that allogeneic MSCs may be both a safe and practical alternative treatment option for equine orthopedic injuries
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