儿童腘窝囊肿液中滑膜干细胞的分离及特性鉴定

Isolation and Characterization of the Synovial-derived Mesenchymal Stem Cells from Popliteal Cyst Fluid in Pediatric Patients

为了从儿童腘窝囊肿液中分离滑膜间充质干细胞（synovium-derived mesenchymal stem cells,SMSCs）,进行体外培养,鉴定其干细胞特性。本实验采用抽取囊肿液、细胞贴壁培养法从腘窝囊肿患儿的囊肿液中分离出滑膜干细胞,进行体外培养、细胞形态学观察、细胞生长曲线绘制及周期的分析,成骨、成脂、成软骨分化能力检测,流式细胞术检测细胞表面标志物,免疫荧光检测干细胞标志物。实验结果显示可采取本方法稳定地从儿童腘窝囊肿液中获取滑膜间充质干细胞,外观梭型状,体外成指数生长,高表达滑膜间充质干细胞表面标记物（CD73,CD90,CD105）,表达胚胎间质细胞标记物波形蛋白（vimentin）和成纤维细胞中α-平滑肌肌动蛋白（α-SMA）,经定向诱导培养基培养,可分化为骨,软骨和脂肪细胞,我们可以确定所分离出的细胞即为滑膜间充质干细胞（SMSCs）。本研究证明可从儿童腘窝囊肿液中获取滑膜间充质干细胞,是间充质干细胞有价值的来源。

To isolate and identify the synovial-derived mesenchymal stem cells from popliteal cyst fluid in pediatric patients. Synovial-derived mesenchymal stem cells were collected from the popliteal cyst fluid in pediatric patients by cyst fluid extraction and adherent culture, and the morphology, growth curve and cell cycle of cells were observed in vitro. The expression of surface markers of cells was analyzed by flow cytometry, and the stem cell markers detected by immunofluorescence. Osteogenic, adipogenic, chondrogenic differentiation and cell secretory factors were assessed to characterize the synovial fluid MSCs. Result showed that we could steadily collect the synovial mesenchymal stem cells from popliteal cyst fluid in pediatric patients, and the cells were shuttle appearance and logarithmic growth, and highly express the synovial mesenchymal stem cells surface markers（CD73, CD90, CD105）, interstitial cell marker（vimentin） and alpha-smooth muscle actin alpha（α-SMA）.These cells can differentiate into bone, cartilage and fat cells by directional inducing medium. We could confirm that the collected cells were synovial mesenchymal stem cells. In conclusion, we can obtain synovial mesenchymal stem cells from the popliteal cyst fluid in pediatric patients, which have a similar morphology and cell phenotype to bone-marrow-derived MSCs, and could be a valuable source of MSCs.