Zayed Mohammed, Caniglia Christopher, Misk Nabil, Dhar Madhu S
Department of Large Animal Clinical Sciences, College of Veterinary Medicine, University of Tennessee, Knoxville, TN, USA; Department of Animal Surgery, College of Veterinary Medicine, South Valley University, Qena, Egypt.
Manor Equine Hospital , Monkton, MD , USA.
Front Vet Sci. 2017 Jan 18;3:121. doi: 10.3389/fvets.2016.00121. eCollection 2016.
Mesenchymal stem cells (MSCs) have been demonstrated to be useful for cartilage tissue regeneration. Bone marrow (BM) and synovial fluid (SF) are promising sources for MSCs to be used in cartilage regeneration. In order to improve the clinical outcomes, it is recommended that prior to clinical use, the cellular properties and, specifically, their chondrogenic potential must be investigated. The purpose of this study is to compare and better understand the chondrogenic potential of equine bone marrow-derived mesenchymal stem cells (BMMSCs) and synovial fluid-derived mesenchymal stem cells (SFMSCs) populated from the same equine donor. BM- and SF-derived MSCs cultures were generated from five equine donors, and the MSCs were evaluated for their morphology, proliferation, trilineage differentiation, and immunophenotyping. Differences in their chondrogenic potentials were further evaluated quantitatively using glycosaminoglycan (GAG) content and immunofluorescence of chondrogenic differentiation protein markers, SRY-type HMG box9, Aggrecan, and collagen II. The BMMSCs and SFMSCs were similar in cellular morphology, viability, and immunophenotype, but, varied in their chondrogenic potential, and expression of the key chondrogenic proteins. The SFMSCs exhibited a significant increase in GAG content compared to the BMMSCs ( < 0.0001) in three donors, suggesting increased levels of chondrogenesis. The expression of the key chondrogenic proteins correlated positively with the GAG content, suggesting that the differentiation process is dependent on the expression of the target proteins in these three donors. Our findings suggest that even though SFMSCs were hypothesized to be more chondrogenic relative to BMMSCs, there was considerable donor-to-donor variation in the primary cultures of MSCs which can significantly affect their downstream application.
间充质干细胞(MSCs)已被证明可用于软骨组织再生。骨髓(BM)和滑液(SF)是用于软骨再生的MSCs的有前景的来源。为了改善临床结果,建议在临床使用前,必须研究细胞特性,特别是它们的软骨生成潜力。本研究的目的是比较并更好地理解来自同一马供体的马骨髓间充质干细胞(BMMSCs)和滑液间充质干细胞(SFMSCs)的软骨生成潜力。从五匹马供体中生成了BM和SF来源的MSCs培养物,并对MSCs的形态、增殖、三系分化和免疫表型进行了评估。使用糖胺聚糖(GAG)含量和软骨分化蛋白标志物SRY型HMG盒9、聚集蛋白聚糖和胶原蛋白II的免疫荧光定量进一步评估它们软骨生成潜力的差异。BMMSCs和SFMSCs在细胞形态、活力和免疫表型上相似,但在软骨生成潜力和关键软骨生成蛋白的表达上有所不同。在三个供体中,与BMMSCs相比,SFMSCs的GAG含量显著增加(<0.0001),表明软骨生成水平增加。关键软骨生成蛋白的表达与GAG含量呈正相关,表明在这三个供体中分化过程依赖于靶蛋白的表达。我们的研究结果表明,尽管相对于BMMSCs,SFMSCs被假设具有更强的软骨生成能力,但在MSCs的原代培养中,供体之间存在相当大的差异,这可能会显著影响它们的下游应用。