Braun Julian, Hack Anita, Weis-Klemm Michaela, Conrad Sabine, Treml Sabrina, Kohler Konrad, Walliser Ulrich, Skutella Thomas, Aicher Wilhelm K
Center for Regenerative Medicine, University Hospital Tübingen, D-72076 Tübingen, Germany.
Am J Vet Res. 2010 Oct;71(10):1228-36. doi: 10.2460/ajvr.71.10.1228.
To evaluate the proliferative behavior, telomere length, immunophenotype, and differentiation capacity of equine adipose tissue-derived mesenchymal stem cells (AT-MSCs).
6 adult racing horses treated for articular Injury but otherwise healthy.
AT-MSCs were Isolated from horses and expanded In Dulbecco modified Eagle medium enriched with fetal bovine serum and antimicrobials. Expression of cell surface antigens and telomere length were Investigated via flow cytometry Differentiation of MSCs Into chondrocytes, osteoblasts, and adipocytes was Induced In vitro by specific stimuli and was evaluated by analyzing marker genes with quantitative reverse transcriptase PCR assays and immunocytochemical and cytologie evaluations.
Equine MSCs could be cultured up to the fifth passage before signs of senescence, apoptosis, and detachment Indicated cellular exhaustion. However, the AT-MSCs from 2 of 6 horses survived to later passages with Increased doubling rates and telomere lengths. The cells had a typical phenotype, with expression of CD14, CD73, CD90, CD105, CD140b, and CD164 antigens and a lack of CD34 and CD45 antigens. The cells also had a strong potential to differentiate Into osteoblasts, as characterized by Intense von Kossa and alizarin red staining as well as high Induction of osteopontin. Chondrogenic differentiation was detected via Alelan blue staining and expression of aggrecan and type II collagen Adipogenesis was Induced in AT-MSCs by supplementation of differentiation media with rabbit serum.
Equine AT-MSCs representa suitable cellular source for regenerative treatment of bone or cartilage defects, particularly when expanded In vitro for only a few passages.
评估马脂肪组织来源间充质干细胞（AT-MSCs）的增殖行为、端粒长度、免疫表型和分化能力。
6匹成年赛马，因关节损伤接受治疗，但其他方面健康。
从马身上分离出AT-MSCs，并在添加胎牛血清和抗菌剂的杜尔贝科改良伊格尔培养基中进行扩增。通过流式细胞术研究细胞表面抗原的表达和端粒长度。通过特定刺激在体外诱导MSCs分化为软骨细胞、成骨细胞和脂肪细胞，并通过定量逆转录酶PCR分析标记基因、免疫细胞化学和细胞学评估进行评价。
马MSCs可培养至第5代，之后衰老、凋亡和脱离迹象表明细胞耗竭。然而，6匹马中有2匹马的AT-MSCs存活至后期传代，倍增率和端粒长度增加。这些细胞具有典型的表型，表达CD14、CD73、CD90、CD105、CD140b和CD164抗原，不表达CD34和CD45抗原。这些细胞也具有很强的分化为成骨细胞的潜力，表现为强烈的冯库萨染色和茜素红染色以及骨桥蛋白的高诱导。通过阿利新蓝染色以及聚集蛋白聚糖和II型胶原的表达检测到软骨形成分化。通过在分化培养基中添加兔血清诱导AT-MSCs发生脂肪生成。
马AT-MSCs是骨或软骨缺损再生治疗的合适细胞来源，特别是在体外仅扩增几代时。
