Evaluation of the osteogenic and chondrogenic differentiation capacities of equine adipose tissue-derived mesenchymal stem cells
Evaluation of the osteogenic and chondrogenic differentiation capacities of equine adipose tissue-derived mesenchymal stem cells
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DOI：
10.2460/ajvr.71.10.1228
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发表时间：
2010-10-01
影响因子：
1
通讯作者：
Aicher, Wilhelm K.
中科院分区：
文献类型：
--
作者：
Braun, Julian;Hack, Anita;Aicher, Wilhelm K.
目的：研究马脂肪组织来源的间充质干细胞（AT-MSCs）的增殖特性、端粒长度、免疫表型和分化能力。通过流式细胞术研究细胞表面抗原的表达和端粒长度。分化成软骨细胞，成骨细胞和脂肪细胞的间充质干细胞诱导在体外由特定的刺激，并通过分析标记基因与定量逆转录酶PCR检测和免疫细胞化学和细胞学evaluations.Results-Equine间充质干细胞可以培养到第五代之前的衰老，凋亡和脱离的迹象表明细胞耗尽。然而，来自6匹马中的2匹的AT-MSC存活到以后的传代，具有增加的倍增率和端粒长度。这些细胞具有典型的表型，表达CD 14、CD 73、CD 90、CD 105、CD 140 b和CD 164抗原，缺乏CD 34和CD 45抗原。细胞也有很强的潜力分化成骨细胞，其特征在于强烈的von Kossa和茜素红染色以及高诱导骨桥蛋白。通过阿辛蓝染色和聚集蛋白聚糖和II型胶原的表达检测软骨分化。脂肪生成诱导AT-MSCs通过补充分化培养基与兔serum.Conclusions和临床相关性马AT-MSCs代表一个合适的细胞来源再生治疗骨或软骨缺损，特别是当在体外扩大只有几个通道。(Am J Vet Res 2010;71：1228-1236）
Objective-To evaluate the proliferative behavior, telomere length, immunophenotype, and differentiation capacity of equine adipose tissue derived mesenchymal stem cells (AT-MSCs).Animals-6 adult racing horses treated for articular injury but otherwise healthy.Procedures-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 cytologic evaluations.Results-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 Alcian blue staining and expression of aggrecan and type II collagen. Adipogenesis was induced in AT-MSCs by supplementation of differentiation media with rabbit serum.Conclusions and Clinical Relevance-Equine AT-MSCs represent a suitable cellular source for regenerative treatment of bone or cartilage defects, particularly when expanded in vitro for only a few passages. (Am J Vet Res 2010;71:1228-1236)