软骨细胞上清液诱导滑膜间充质干细胞微团培养成软骨

背景：滑膜间充质干细胞在体外具有多向分化的能力，有望成为软骨组织工程中治疗软骨缺损的种子细胞，在其向软骨细胞分化过程中，合适的生长因子起了重要作用。目的：利用富含生长因子的软骨细胞上清液诱导滑膜间充质干细胞向软骨细胞分化，并对其鉴定。方法：采用消化法分别获得SD大鼠滑膜间充质干细胞、软骨细胞。收集软骨细胞上清液离心、过滤冻存备用。培养滑膜间充质干细胞至第3代后离心成微团，并用软骨细胞上清液进行成软骨诱导分化，通过形态学观察、免疫组织化学法、RT-PCR检测进行鉴定。结果与结论：滑膜间充质干细胞使用软骨细胞上清液成软骨诱导21 d后，微团可见似软骨样组织。免疫组化法进行Ⅱ型胶原鉴定，基质能被Ⅱ型胶原染色，细胞染色呈现棕黄色。RT-PCR结果显示诱导后的微团表达软骨特异性基因Ⅱ型胶原和蛋白聚糖。证实软骨细胞分泌的可溶性因子可以诱导大鼠滑膜间充质干细胞向软骨方向分化。中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程全文链接：

Chondrocyte supernatant induces chondrogenesis and pellet cultivation of rat synovial mesenchymal stem cells

BACKGROUND: Synovial mesenchymal stem cells have the ability of multilineage differentiation in vitro, which are expected to be seed cells for the treatment of cartilage defects in cartilage tissue engineering. Appropriate growth factors are critical for the chondrocyte differentiation of synovial mesenchymal stem cells.OBJECTIVE:To study the role of secreted factors by chondrocytes to induce chondrogenesis of synovial mesenchymal stem cells. METHODS:The synovial mesenchymal stem cells and chondrocytes were harvested from rat knee joints and cultured through the digestion method. The supernatant was collected from chondrocytes, and centrifuged, filtered and cryopreserved. The third passage synovial mesenchymal stem cells centrifuged as pellets were cultured in the chondrocyte supernatant for 21 days. And the cells morphology was examined and the type II collagen and aggrecan were detected through immunohistochemistry and RT-PCR. RESULTS AND CONCLUSION:The synovial mesenchymal stem cell pellets cultured in the chondrocyte supernatant became cartilage-like tissue after 21 days. The type II collagen was detected positively in the matrix of synovial mesenchymal stem cell pellet immunohistochemically. RT-PCR examination showed that the type II collagen and aggrecan expressed in the synovial mesenchymal stem cellpellet cultured in the chondrocyte supernatant. It suggested that synovial mesenchymal stem cell could be induced to differentiate into chondrocytes depending on soluble factors secreted by chondrocytes.