兔骨髓间充质干细胞的制备及诱导成骨的实验研究.pdfVIP

第13卷4期 天 津 医科 大 学 学 报 Vo1．13．No 4 2007年12月 JOURNAL OF TIANJIN MEDICAL UNIVERSITY Dec．2007 507 兔骨髓问充质干细胞的制备及诱导成骨的实验研究 潘 丽，李晓眠，张黎龙，李晶哲 (天津医科大学微生物学教研室，天津300070) [摘要]目的：在体外原代培养兔骨髓间充质干细胞，观察兔骨髓间充质干细胞的体外生长特性，为用于组织工程学的 种子细胞培养提供实验基础。方法：自兔股骨、胫骨取骨髓．分离骨髓问充质干细胞，分别采用常规培养液及含矿化液 的培养液两组进行培养，对第3代细胞进行酶动力学法碱性磷酸酶(ALP)活性检测及钙结节茜素红染色．观察细胞在 体外长期培养时钙结节形成情况。结果：两组细胞其培养液中的AIJP含量不断升高，经过两独立样本t检验．矿化液 诱导组第2，6，10天的AIJP水平较正常对照组均明显升高(P0．05)。细胞表现出成骨细胞特性，在体外长期不传代 培养时可形成钙结节。结论：兔骨髓问充质于细胞在体外适当的培养条件下。增殖能力很强，可以向成骨细胞方向分 化．可作为组织工程学的种子细胞。 [关键词]骨髓间充质干细胞；原代培养；兔：碱性磷酸酶；组织工程 [中图分类号]Q813．1 [文献标识码]A [文章编号]1006～8147(2007)04—0507—04 Study on induced bone formation of rabbit marrow mesenchymal stem cells PAN Li，LI Xiao—mian，ZHANG Li-long，LI Jing-zhe (Department of Microbiology，Tianjin Medical University，Tianjin 300070，China) ABSTRACT Objective：To establish primary bone marrow mesenchymal stem cells(BMSCs)of rabbit in vitro and observe its growth characteristics，SO as to provide experimental framework to cultivate various seed cells for tissue engineering．Methods：Bone marrow was extracted from the femoral and shin bone 0f rabbit，BMSCs were separated from bone marrow and cultivated in two groups，that is DMEM media group and DMEM with mineralized liquid group respectively．To monitor the activity of alkaline phosphatase(ALP1 of passage 3 BMSCs by means of enzyme kinetics and to observe BMSC——derived calcification nodule by staining with alizarin bordeaux．Results：ALP level in both groups increased gradually．Independent samples t test showed the ALP activity in the DMEM with mineralized liquid group on the 2nd，6th，1 0th day was significantly higher than that in DMEM media group 0．o5)．BMSCs had the