SD大鼠胚胎后肾间充质干细胞的分离培养及生物学特性研究

Isolation and characterization of SD rat embryonic kidney metanephric mesenchyme stems cells

目的对SD大鼠胚胎后肾间充质细胞(MMSCs)进行分离培养,并检测其生物学特性。方法采用胚胎后肾微组织块接种培养法,检测其生长曲线和生长周期,并进行细胞表面抗原和成骨能力的鉴定。结果MMSCs体外培养生长状况良好,具有活跃的增殖能力。细胞周期显示90%以上细胞处于G0/G1期。表面抗原检测绝大多数细胞阳性表达波形蛋白(vimen-tin)和纤维连接蛋白(fibronectin),几乎不表达CD34。在体外能够向成骨细胞分化。结论原代分离、培养的细胞为MMSCs,细胞纯度高、扩增迅速、生物学性状稳定,可为组织工程提供比较理想的种子细胞。

Obgective To isolate and cultivate metanephric mesenchyme stem cells（MMSCs） primarily derived from metanephric tissue of embryonic rats in vitro and identify their biological characteristics. Methods Pieces of rat metanephric tissue were explan- ted into plastic flask , and cells were grown in the tissue pieces. The cell growth curve and cell cycle were measured and surface anti- gens were detected by immunohistochemistry. Osteoblast differentiation was studied by Alizarin red staining. Results The cells showed shuttle shape with single nuclear and strong proliferative ability in vitro. Cultured cells could be propagated to 10 passages and remained the phenotype of mesenchymal cells. The cell cycle analysis showed that more than 90~ of BMSCs were at Go/G1 phase. Immunochemistry staining revealed that the cells were positive for vimentin and fibronectin,but negative for CD34. The cells could be induced to differentiate into osteocytes under special conditions. Conclusion MMSCs obtained from SD rats embryonic kidney grew well in vitro and had high purity and proliferation activity. The biological properties was stabilization.