SD大鼠间充质干细胞的分离培养及生物学特性研究

Isolating culture and biological characteristics of mesenchymal stem cells in SD rats

目的对SD大鼠不同组织的间充质干细胞(MSCs)进行分离培养并检测其生物学特性。方法分离SD大鼠骨髓、骨密质来源的MSCs,进行细胞表面抗原和细胞生长周期的鉴定。结果两种组织来源的细胞都呈成纤维细胞样,单个核,增殖能力强。细胞免疫组织化学结果显示,培养的细胞表达波形蛋白(vimention)、纤维连接蛋白(fibronectin)但不表达角蛋白(keratin)。细胞可传代10代以上并保持细胞表型。细胞周期显示90%以上细胞处于细胞静止期。结论分离、培养的两种组织的细胞均为MSCs,在体外培养条件下增殖能力强、表型稳定,可作为组织工程的种子细胞。

Objective To isolate and cultivate mesenchymal stem cells （MSCs） in different tissues of SD rats, and investigate their biological characteristics. Methods MSCs were obtained from bone marrow and compact bone of SD rats and cultivated in DMEM F12 medium. MSCs membrane antigens were identified with immunocytochemical method. The cell cycle was measured by flow cytome try. Results The cells showed shuttle shape with single nuclear and strong proliferative ability in vitro. Immunochemistry staining assay revealed significant expression of vimention, fibronectin, but no keratin expression was found. The cultured cells could be propagated to 10 generations and remained the phenotype of mesenchymal cells. The cell cycle analysis showed that more than 90% of MSCs were in G0/G1 phase. Conclusion Highly purified MSCs obtained from bone marrow and compact bone of SD rats can be propagated and have active proliferative ability and phenotype stability. The MSCs can be used as seeding cells in tissue engineering.