大鼠后肾间充质细胞的体外培养  

Culture of metanephric mesenchymal cells derived from metanephric tissue of embryonic rat in vitro

作　　者：周剑锋[1] 袁发焕[1] 李娜 侯卫平[1] 

机构地区：[1]第三军医大学附属新桥医院肾内科全军肾脏病中心,重庆400037 [2]解放军第一零七中心医院肾内科

出　　处：《中华肾脏病杂志》2007年第10期668-672,共5页Chinese Journal of Nephrology

基　　金：重庆市自然科学基金（2006BB5055）

摘　　要：目的建立大鼠后肾间充质细胞（MMC）的培养方法,为体外研究肾脏纤维化的发病机制和防治提供细胞技术平台。方法采用胚胎后肾组织块接种培养法。通过相差显微镜观察培养的细胞形态。5-溴脱氧尿嘧啶核苷（BrdU）法及四唑盐比色（MTF）法测定细胞增殖。流式细胞术及细胞免疫化学染色法检测细胞表型标记蛋白以及成脂、成骨多向诱导分化等方法鉴定。结果培养的细胞呈梭形成纤维细胞样,单个核,核质比大,增殖能力强。流式细胞术显示细胞不表达CD31、CD34、CD45,但明显表达CD29、CD166。细胞免疫化学结果显示该细胞不表达E-钙黏蛋白（E-cadherin）,但明显表达波形蛋白（vimentin）和纤连蛋白（fibronectin）。细胞可以传代至10代以上并且保持其间充质细胞表型,且在成脂及成骨诱导培养的条件下具有多向分化的能力。结论原代培养的细胞为间充质细胞并且可以传代,具备较强的细胞增殖能力和表型稳定性,为后续实验提供了可靠资源。Objective To establish a method culturing metanephric mesenchymal ceils （MMCs） primarily derived from metanephric tissue of embryonic rats in vitro. Methods Pieces of rat metanephric tissue were explanted into plastic flask, and cells were grown in the tissue pieces. Cells were identified by inverted phase contrast microscope. Phenotypic protein examination was performed via flow cytometry and immunochemistry staining. Cells proliferation was measured by MTT and BrdU, and multi-direction differentiation was also induced. Results The cells showed shuttle shape with single nuclear in each one. Flow cytometry and immunochemistry staining revealed that significant expression of vimentin, fibronectin, CD29 and CD166 was positive over all of the cells and expression of CD31, CD34, CD45 and E-cadherin was negative in contrast, which confirmed the mesenchymal cell phenotype. Cultured cells could be propagated to 10 passages and remained the phenotype of mesenchymal cells. The cells could be induced to differentiate to adipocytes and osteocytes under special conditions. Conclusion Cultured mesenchymal cells from metanephric tissue of embryonic rats can be propagated and have strong proliferation ability and stability.

关 键 词：后肾 间充质细胞 原代培养 

分 类 号：R686[医药卫生—骨科学]