Prado Aline Ambrogi Franco, Favaron Phelipe Oliveira, da Silva Luis Claudio Lopes Correia, Baccarin Raquel Yvonne Arantes, Miglino Maria Angelica, Maria Durvanei Augusto
Department of Surgery, School of Veterinary Medicine and Animal Science, University of Sao Paulo, Av. Prof. Dr. Orlando Marques de Paiva, 87, Cidade Universitária, 05508-270, São Paulo, SP, Brazil.
Laboratory of Biochemical and Biophisic, Butantan Institute, Av. Dr. Vital Brasil, 1500, 05503-900, Sao Paulo, Brazil.
BMC Vet Res. 2015 Nov 10;11:281. doi: 10.1186/s12917-015-0531-5.
Isolation of mesenchymal stem cells (MSCs) in equines, has been reported for different tissues including bone marrow, adipose, umbilical cord, peripheral blood, and yolk sac. In regard to the MSCs derived from synovial fluid (SF) or membrane (SM), there is data available for humans, dogs, pigs, goats and horses. Especially in equines, these cells have being considered promising candidates for articular regeneration. Herein, we established and characterized MSCs obtained from equine SF and SM. Samples were obtained during arthroscopy and cultured using MEM (Minimum Essential Medium). MSCs were characterized by morphology and expression of specific markers for stem cells, pluripotency, inflammation, and cell cycle.
The medium MEM was more effective (97% ± 2) to maintain both cultures. The cultures were composed by adherent cells with fibroblast-like shape, which had a growth pattern represented by a sigmoidal curve. After the expansion, the cells were analyzed by flow cytometry for stem cells, inflammatory, and cell cycle markers, and both lineages showed significant expression of CD45, Oct3/4, Nanog, CD105, CD90, CD34, CD117, CD133, TRA-1-81, VEGF, and LY6a. In contrast, there were differences in the cell cycle phases between the lineages, which was not observed in relation to the mitochondrial electrical potential.
Given the large impact that joint pathology has on the athletic performance horses, our results suggested that the SF and SM are promising sources of stem cells with satisfactory characteristics of growth and gene expression that can be used in equine regenerative medicine.
已报道从包括骨髓、脂肪、脐带、外周血和卵黄囊在内的不同组织中分离马的间充质干细胞(MSCs)。关于源自滑液(SF)或滑膜(SM)的MSCs,已有针对人类、犬、猪、山羊和马的数据。特别是在马中,这些细胞被认为是关节再生的有前景的候选细胞。在此,我们建立并鉴定了从马的SF和SM中获得的MSCs。样本在关节镜检查期间获取,并使用MEM(最低限度基本培养基)进行培养。通过形态学以及干细胞、多能性、炎症和细胞周期的特定标志物的表达对MSCs进行鉴定。
培养基MEM在维持两种培养物方面更有效(97%±2)。培养物由具有成纤维细胞样形状的贴壁细胞组成,其生长模式呈S形曲线。扩增后,通过流式细胞术分析细胞的干细胞、炎症和细胞周期标志物,两个谱系均显示CD45、Oct3/4、Nanog、CD105、CD90、CD34、CD117、CD133、TRA-1-81、VEGF和LY6a有显著表达。相比之下,两个谱系在细胞周期阶段存在差异,而在线粒体电位方面未观察到差异。
鉴于关节病理学对赛马运动表现有很大影响,我们的结果表明,SF和SM是有前景的干细胞来源,具有令人满意的生长和基因表达特征,可用于马的再生医学。