Characterization and profiling of immunomodulatory genes of equine mesenchymal stromal cells from non-invasive sources.
Abstract: Mesenchymal 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. Methods: Equine 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. Results: While 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). Conclusions: Whereas 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.
Publication Date: 2014-01-13 PubMed ID: 24418262PubMed Central: PMC4055120DOI: 10.1186/scrt395Google Scholar: Lookup The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Journal Article
Summary
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The research investigates the properties of equine Mesenchymal stromal cells (MSCs) as isolated from non-invasive sources such as umbilical cord blood (UCB), umbilical cord matrix (UCM) and equine peripheral blood (PB). It finds that UCB and PB are promising sources for these cells, while UCM was less successful due to contamination risks.
Research Methods
- The MSCs were collected from three non-invasive sources and from six individual mares, minimizing variability by matching samples from the same mare and for UCB and UCM, the same foal.
- The researchers analyzed the success rate of isolation, proliferation capacity, tri-lineage differentiation ability, immunophenotypic protein, and immunomodulatory mRNA profiles of these MSCs.
- Linear regression models were used to determine the association between the source of MSCs and a variety of factors, such as the moment of first observation, the moment of first passage, cell proliferation data, the expression of markers related to cell immunogenicity, and the mRNA profile of immunomodulatory factors.
- Hepatocyte growth factor (HGF) was excluded as no normal distribution could be obtained. The non-parametric Kruskal-Wallis test was used instead to evaluate the association between the MSC source and HGF mRNA expression.
Findings
- The study found that MSCs could be successfully isolated from UCB and PB samples, but not from UCM due to contamination issues.
- The researchers found that UCB-derived MSCs proliferated significantly quicker than PB or UCM-derived MSCs.
- Both UCB and PB-derived MSCs demonstrated the ability to be differentiated toward the osteo-, chondro-, and adipogenic lineage, whereas UCM-derived MSCs only showed potential for chondro- and adipogenic differentiation.
- Irrespective of the source, the MSCs expressed immunomodulatory genes such as CD40, CD80, HGF, and TGFβ. However, no mRNA expression was found for CD86, IDO, and TNFα.
Conclusions
- The research concluded that although UCM was less viable due to high contamination risks and low success rates, UCB and PB emerged as promising alternative sources of MSCs, particularly considering the use of allogenic MSCs for future therapeutical applications.
Cite This Article
APA
De Schauwer C, Goossens K, Piepers S, Hoogewijs MK, Govaere JL, Smits K, Meyer E, Van Soom A, Van de Walle GR.
(2014).
Characterization and profiling of immunomodulatory genes of equine mesenchymal stromal cells from non-invasive sources.
Stem Cell Res Ther, 5(1), 6.
https://doi.org/10.1186/scrt395 Publication
Researcher Affiliations
MeSH Terms
- Adipogenesis
- Animals
- B7-1 Antigen / genetics
- B7-1 Antigen / metabolism
- B7-2 Antigen / genetics
- B7-2 Antigen / metabolism
- CD40 Antigens / genetics
- CD40 Antigens / metabolism
- Cells, Cultured
- Female
- Fetal Blood / cytology
- Gene Expression Profiling
- Hepatocyte Growth Factor / genetics
- Hepatocyte Growth Factor / metabolism
- Horses
- Indoleamine-Pyrrole 2,3,-Dioxygenase / genetics
- Indoleamine-Pyrrole 2,3,-Dioxygenase / metabolism
- Mesenchymal Stem Cells / cytology
- Mesenchymal Stem Cells / immunology
- Mesenchymal Stem Cells / metabolism
- Osteogenesis
- RNA, Messenger / genetics
- RNA, Messenger / metabolism
- Transforming Growth Factor beta / genetics
- Transforming Growth Factor beta / metabolism
- Tumor Necrosis Factor-alpha / genetics
- Tumor Necrosis Factor-alpha / metabolism
- Umbilical Cord / cytology
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