大鼠跟腱来源肌腱干细胞的分离培养及鉴定  

Isolation and identification of tendon stem cells from rat Achilles's tendon

作　　者：胡超[1] 唐康来[1] 陈万[1] 马林[1] 杨广华[1] 

机构地区：[1]第三军医大学西南医院骨科,全军矫形外科中心,重庆400038

出　　处：《第三军医大学学报》2013年第11期1097-1101,共5页Journal of Third Military Medical University

基　　金：国家自然科学基金(81071464)~~

摘　　要：目的从大鼠跟腱标本中分离培养肌腱干细胞(tendon stem/progenitor cells,TSCs)并加以鉴定。方法大鼠跟腱经消化后,获取的有核细胞按不同密度在37℃,5%CO2条件下培养,原代为P0代,P1～P3代用于实验研究。观察其克隆形成和增殖能力,用流式细胞仪和免疫组化染色方法对其干细胞标志物予以鉴定,以不同的全成分诱导分化液诱导,观察其多向分化潜能。结果大鼠跟腱来源肌腱干细胞具有克隆生长特性,表面标记物表达为CD44h(+)、CD90(+)、CD34(-)、CD106(-),免疫荧光染色显示SSEA4、Nucleostemin及Ⅰ型胶原蛋白表达均为阳性。经诱导后能够向成骨、成脂肪、成软骨多向分化。结论大鼠跟腱来源肌腱干细胞具有干细胞特性,不同条件下可能向不同方向分化。Objective To isolation and identify tendon stem cells from rat Achilles’s tendon. Methods Nucleated cells isolated from rat Achilles’s tendon tissues after collagenase digestion were plated at different cell densities and cultured at 37 ℃, 5%CO2. These cells were at passage 0 （P0）, and then the cells from passages 1-3 （P1-P3） were used for the following experiments. Cell proliferation assay and colony assay were used to observe the self-renewal and proliferative potential of the obtained cells. Immunocytochemical staining and flow cytometry were employed to detect the expression of stem cell markers and lineage-specific markers. Osteogenic, chondrogenic, and adipogenic multi-differentiation potentials were observed by cultured in different medium. Results These cells were CD44h（＋）, CD90（＋）, CD34（-）, and CD106（-） indicated that they shared some common properties with mesenchymal stem cells. They also exhibited their unique characteristics by expressing tenogenic and stem cell markers SSEA4, nucleostemin, anti-CollagenⅠ, but not expressing CollagenⅢ. The cells also had osteogenic, chondrogenic, and adipogenic differentiation potentials by cultured in different conditions. Conclusion Tendon-derived stem cells with multi-differentiation potentials are successfully isolated from rat Achilles’s tendon under the optimized growth and differentiation conditions.

关 键 词：跟腱 肌腱干细胞 分离 鉴定 大鼠 

分 类 号：R322.73[医药卫生—人体解剖和组织胚胎学] R329-33[医药卫生—基础医学]