滑膜间充质干细胞分离纯化后的生物学特性

背景：大量的研究报道证明滑膜间充质干细胞在细胞形态、免疫表型、集落形成能力和分化潜能等方面与骨髓间充质干细胞相似，但在向软骨分化的能力上，滑膜间充质干细胞明显优于骨髓间充质干细胞。目的：探讨滑膜间充质干细胞作为半月板软骨组织工程种子细胞的可行性。方法：通过有限稀释单克隆培养法将滑膜间充质干细胞从兔滑膜组织中分离出来并加以纯化，在体外培养条件下对其形态学、超微结构、分子表型、增殖动力学、核型以及致瘤性等进行分析。结果与结论：从兔滑膜细胞中分离纯化出滑膜间充质干细胞，体外单层培养具有极强的增殖能力，在第6天时达到生长的最高峰，倍增时间为（30．2±2．4）h。流式细胞术检测滑膜间充质干细胞表达间充质干细胞一些分子标记CD44、CD90。DNA含量检测、染色体核型分析、荷瘤实验结果表明，分离纯化的滑膜间充质干细胞是正常的二倍体细胞，无致瘤性，因此可作为半月板组织工程的种子细胞。

Biological characteristics of synovial mesenchymal stem cells after isolation and purification

BACKGROUND： A large amount of studies have confirmed that synovial mesenchymal stem ceils have the similarity in cell morphology, immune phenotype, colony forming ability and differentiation potential with bone marrow mesenchymal stem cells. But bone marrow mesenchymal stem cells are better than synovial mesenchymal stem cells in the ability to differentiate into cartilages. OBJECTIVE： To discuss the possibility of using synovial mesenchymal stem cells as seed cells for meniscal tissue engineering. METHODS： The synovial mesenchymal stem cells were isolated from rabbit synovial tissues with limiting dilution mono.clonal culture method, and then the cells were purified. The morphology, ultrastructure, molecular phenotype, proliferation kinetics, karyotype and tumorigenicity of the in vitro cultured cells were analyzed. RESULTS AND CONCLUSION： The synovial mesenchymal stem cells isolated from the rabbit synovial cells had high proliferation capacity during in vitro monolayer culture. The synovial mesenchymal stem cells grew to peak at 6 days, and the doubling time was （30.2±2.4） hours. Flow cytometry results showed the synovial mesenchymal stem cells could express some molecular makers of mesenchymal stem cells, Such as CD44 and CD90. DNA contents check, karyotype test and oncogenicity test confirmed isolated and purified synovial mesenchymal stem cells were the normal diploid cells without tumorigenicity, so the cells can be used as seed cells for meniscal tissue engineering.