Division of Physiology, Department of Basic Sciences, Faculty of Veterinary Medicine, Ferdowsi University of Mashhad, Mashhad, Iran.
Division of Physiology, Department of Basic Sciences, Faculty of Veterinary Medicine, Ferdowsi University of Mashhad, Mashhad, Iran; Stem Cell Biology and Alternative Regenerative Medicine Group, Institute of Biotechnology, Ferdowsi University of Mashhad, Mashhad, Iran.
Res Vet Sci. 2019 Aug;125:235-243. doi: 10.1016/j.rvsc.2019.06.015. Epub 2019 Jul 3.
Equine adipose-derived mesenchymal stem cells (eq-ASCs) possess excellent regeneration potential especially for treatment of musculoskeletal disorders. Besides their common characteristics, MSCs harvested from different species reveal some species-specific and donor-dependent behaviors. Hence, the molecular analysis of MSCs may shed more light on their future clinical application of these cells. This study aimed to investigate some behavioral aspects of eq-ASCs in vitro which may influence the efficacy of stem cell therapy. For this purpose, MSCs of a donor horse were isolated, characterized and expanded under normal culture conditions. During continuous culture condition, eq-ASCs were started to formed aggregated structures that was accompanied with the up-regulation of migratory related genes including transforming growth factor beta 1 (TGFB1) and its receptor 3 (TGFBR3), and snail family transcriptional repressor 1 (SNAI1), E-cadherin (CDH1) and β-catenin (CTNNB1). Moreover, the expression of a musculoskeletal progenitor marker, scleraxis bHLH transcription factor (SCX), was also increased after 3 days. In order to clarify the impact of TGFB signaling pathway on cultured cells, gain- and loss-of-function treatment by TGFB3 and SB431542 (TGFB inhibitor) were performed, respectively. We found that TGFB3 treatment exaggerated the aggregate formation effects, in some extend via induction of cytoskeletal actin rearrangement, while inhibition of TGFB signaling pathway by SB431542 reversed this phenomenon. Overall, our findings support the fact that eq-ASCs have an inherent capacity for migration, which was enhanced by TGFB3 treatment and, this ability may play crucial role in cell motility and wound healing of transplanted cells.
马脂肪间充质干细胞（eq-ASCs）具有出色的再生潜力，尤其适用于治疗肌肉骨骼疾病。除了它们的共同特征外，从不同物种中分离出的间充质干细胞表现出一些物种特异性和供体依赖性的行为。因此，间充质干细胞的分子分析可能会更深入地了解这些细胞未来的临床应用。本研究旨在研究 eq-ASCs 在体外的一些行为特征，这些特征可能会影响干细胞治疗的效果。为此，从一匹供体马中分离、鉴定和扩增了间充质干细胞。在连续培养条件下，eq-ASCs 开始形成聚集结构，同时迁移相关基因（包括转化生长因子β 1（TGFB1）及其受体 3（TGFBR3）、Snail 家族转录抑制因子 1（SNAI1）、E-钙黏蛋白（CDH1）和β-连环蛋白（CTNNB1））的表达上调。此外，在第 3 天，肌肉骨骼祖细胞标记物 Scleraxis bHLH 转录因子（SCX）的表达也增加了。为了阐明 TGFB 信号通路对培养细胞的影响，分别进行了 TGFB3 和 SB431542（TGFB 抑制剂）的增益和失活处理。我们发现，TGFB3 处理通过诱导细胞骨架肌动蛋白重排，夸大了聚集形成的作用，而 SB431542 抑制 TGFB 信号通路则逆转了这一现象。总之，我们的研究结果支持这样一个事实，即 eq-ASCs 具有固有迁移能力，TGFB3 处理增强了这种能力，而这种能力可能在移植细胞的细胞迁移和伤口愈合中发挥关键作用。
