Evaluation of equine peripheral blood apheresis product, bone marrow, and adipose tissue as sources of mesenchymal stem cells and their differentation potential

Ahern, Benjamin J.; Schaer, Thomas P.; Terkhorn, Shawn P.; Jackson, Karen V.; Mason, Nicola J.; Hankenson, Kurt D.

doi:10.2460/ajvr.72.1.127

Cited by 25 publications

(24 citation statements)

References 22 publications

(50 reference statements)

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“…To minimalize animal-dependent influences between the different sources, the samples of the three tissue sources were at least matched within the same mare and for UCB and UCM even within the same foal from that specific mare. As such, we significantly reduced the large variability in age and breed of the horses [32]. As an overall conclusion, we propose UCB as the most valuable, non-invasive source for equine MSCs on the basis of the following observations.…”

Section: Discussionmentioning

confidence: 78%

Characterization and profiling of immunomodulatory genes of equine mesenchymal stromal cells from non-invasive sources

et al. 2014

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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.

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Section: Discussionmentioning

confidence: 78%

Characterization and profiling of immunomodulatory genes of equine mesenchymal stromal cells from non-invasive sources

et al. 2014

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“…Several studies reported the characterization of AdMSCs in horses, where the most common processes are osteogenic, adipogenic and chondrogenic (tri-lineage) differentiation of progenitor cells (Vidal et al, 2007(Vidal et al, , 2008Kisiday et al, 2008;Mambelli et al, 2009;Colleoni et al, 2009;Toupadakis et al, 2010;Ahern et al, 2011;Schwarz et al, 2011a,b). Only five recent studies have reported the analyses of surface antigens of AdMSCs by flow cytometry (de Mattos Carvalho et al, 2009;Braun et al, 2010;Pascucci et al, 2011;Raabe et al, 2011;Ranera et al, 2011).…”

Section: mentioning

confidence: 99%

Characterization of mesenchymal stem cells derived from equine adipose tissue

Carvalho

Yamada

Golim

et al. 2013

Arq. Bras. Med. Vet. Zootec.

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Stem cell therapy has shown promising results in tendinitis and osteoarthritis in equine medicine. The purpose of this work was to characterize the adipose-derived mesenchymal stem cells (AdMSCs) in horses through (1) the assessment of the capacity of progenitor cells to perform adipogenic, osteogenic and chondrogenic differentiation; and (2) flow cytometry analysis using the stemness related markers: CD44, CD90, CD105 and MHC Class II. Five mixed-breed horses, aged 2-4 years-old were used to collect adipose tissue from the base of the tail. After isolation and culture of AdMSCs, immunophenotypic characterization was performed through flow cytometry. There was a high expression of CD44, CD90 and CD105, and no expression of MHC Class II markers. The tri-lineage differentiation was confirmed by specific staining: adipogenic (Oil Red O), osteogenic (Alizarin Red), and chondrogenic (Alcian Blue). The equine AdMSCs are a promising type of adult progenitor cell for tissue engineering in veterinary medicine.

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“…However, due to the low concentration of progenitor cells in the mononuclear cellular fraction of peripheral blood and because these cells are very fragile and have a limited capacity for expansion, further studies are required to increase the cell yields and make therapeutic application viable (Koerner et al 2006). Ahern et al (2011) have unsuccessfully attempted to isolate and grow equine peripheral blood-derived progenitor cells by processing blood using an apheresis machine, confirming the difficulty in isolating and cultivating these cells.…”

Section: Introductionmentioning

confidence: 99%

“…The advantage of using PbMSCs is that collecting this material is safer and less invasive than bone marrow puncture (Ahern et al 2011). However, due to the low concentration of progenitor cells in the mononuclear cellular fraction of peripheral blood and because these cells are very fragile and have a limited capacity for expansion, further studies are required to increase the cell yields and make therapeutic application viable (Koerner et al 2006).…”

Section: Introductionmentioning

confidence: 99%

Isolation and characterization of equine peripheral blood-derived multipotent mesenchymal stromal cells

Carvalho

Yamada

Martins³

et al. 2013

Pesq. Vet. Bras.

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The objective of the study was to isolate, cultivate and characterize equine peripheral blood-derived multipotent mesenchymal stromal cells (PbMSCs). Peripheral blood was collected, followed by the isolation of mononuclear cells using density gradient reagents, and the cultivation of adherent cells. Monoclonal mouse anti-horse CD13, mouse anti-horse CD44, and mouse anti-rat CD90 antibodies were used for the immunophenotypic characterization of the surface of the PbMSCs. These cells were also cultured in specific media for adipogenic and chondrogenic differentiation. There was no expression of the CD13 marker, but CD44 and CD90 were expressed in all of the passages tested. After 14 days of cell differentiation into adipocytes, lipid droplets were observed upon Oil Red O (ORO) staining. Twenty-one days after chondrogenic differentiation, the cells were stained with Alcian Blue. Although the technique for the isolation of these cells requires improvement, the present study demonstrates the partial characterization of PbMSCs, classifying them as a promising type of progenitor cells for use in equine cell therapy.

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Evaluation of equine peripheral blood apheresis product, bone marrow, and adipose tissue as sources of mesenchymal stem cells and their differentation potential

Cited by 25 publications

References 22 publications

Characterization and profiling of immunomodulatory genes of equine mesenchymal stromal cells from non-invasive sources

Characterization and profiling of immunomodulatory genes of equine mesenchymal stromal cells from non-invasive sources

Characterization of mesenchymal stem cells derived from equine adipose tissue

Isolation and characterization of equine peripheral blood-derived multipotent mesenchymal stromal cells

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