Trachsel Dagmar S, Stage Hannah J, Rausch Sebastian, Trappe Susanne, Söllig Katharina, Sponder Gerhard, Merle Roswitha, Aschenbach Jörg R, Gehlen Heidrun
Equine Clinic: Surgery and Radiology, Department of Veterinary Medicine, Freie Universität Berlin, Oertzenweg 19b, 14163 Berlin, Germany.
Clinical Unit of Equine Internal Medicine, Department for Companion Animals and Horses, University of Veterinary Medicine Vienna, Veterinärplatz 1, 1210 Vienna, Austria.
Animals (Basel). 2022 Aug 11;12(16):2049. doi: 10.3390/ani12162049.
Physiological particularities of the equine heart justify the development of an in vitro model suitable for investigations of the species-specific equine cardiac electrophysiology. Adipose tissue-derived stromal/stem cells (ASCs) could be a promising starting point from which to develop such a cardiomyocyte (CM)-like cell model. Therefore, we compared abdominal, retrobulbar, and subcutaneous adipose tissue as sources for the isolation of ASCs applying two isolation methods: the collagenase digestion and direct explant culture. Abdominal adipose tissue was most suitable for the isolation of ASCs and both isolation methods resulted in comparable yields of CD45-/CD34-negative cells expressing the mesenchymal stem cell markers CD29, CD44, and CD90, as well as pluripotency markers, as determined by flow cytometry and real-time quantitative PCR. However, exposure of equine ASCs to 5-azacytidine (5-AZA), reportedly inducing CM differentiation from rats, rabbits, and human ASCs, was not successful in our study. More precisely, neither the early differentiation markers and , nor the late CM differentiation markers , and were upregulated in equine ASCs exposed to 10 µM 5-AZA for 48 h. Hence, further work focusing on the optimal conditions for CM differentiation of equine stem cells derived from adipose tissue, as well as possibly from other origins, are needed.
马心脏的生理特性证明了开发一种适用于研究马种特异性心脏电生理学的体外模型的合理性。脂肪组织来源的基质/干细胞（ASCs）可能是开发这种心肌样细胞模型的一个有前景的起点。因此，我们比较了腹部、球后和皮下脂肪组织作为分离ASCs的来源，应用了两种分离方法：胶原酶消化法和直接外植体培养法。腹部脂肪组织最适合分离ASCs，两种分离方法均产生了相当数量的表达间充质干细胞标志物CD29、CD44和CD90以及多能性标志物的CD45-/CD34阴性细胞，这通过流式细胞术和实时定量PCR确定。然而，在我们的研究中，将马ASCs暴露于据报道可诱导大鼠、兔子和人ASCs向心肌细胞分化的5-氮杂胞苷（5-AZA）并不成功。更确切地说，在暴露于10 µM 5-AZA 48小时的马ASCs中，早期分化标志物和，以及晚期心肌细胞分化标志物、和均未上调。因此，需要进一步开展工作，重点关注源自脂肪组织以及可能源自其他来源的马干细胞向心肌细胞分化的最佳条件。
