改良组织块法分离培养兔成纤维样滑膜细胞

目的通过改良组织块细胞培养法,建立一种简单,可行的兔成纤维样滑膜细胞体外培养方法。方法无菌条件下取正常兔膝关节滑膜组织,剔净后剪成碎块,用灭菌滤纸吸掉组织上附着的水分,碎片接种于培养皿,观察细胞生长状况及形态特征。取第3代对数期细胞,用CCK-8法绘制其生长曲线,并用免疫荧光法检测波形蛋白的表达情况。结果体外培养的兔成纤维样滑膜细胞形态为长梭形纤维样,细胞生长力较旺盛,生长曲线为典型的"S"型,免疫荧光检测显示第3代细胞强表达波形蛋白。结论改良组织块法可以分离培养出兔成纤样维滑膜细胞,方法简便,成功率高,满足实验要求。

Primary Culture of Rabbit Fibroblast-like Synoviocytes using the Improved explant culture method

Abstrat] Objective: To establish an easier and better primary culter technique suitable to rabbit fibroblast-like synoviocytes(FLS) by modified tissue cell culture method. Method: Healthy rabbit joint synovial layers were cut into small pieces and siphon off water attached on the organization with sterile filter paper and then cultured. The growth status and morphology were observed. The growth curve of the 3rd passage cells was measured by CCK-8 assay, and the expression of vimentin was tested by immunocytochemistry. Results: The FLS cultured in vitro exhibited a spindle-shaped appearance and could rapidly expand. The cell growth curve was typical of S type, and the cells highly expressed vimentin. Conclusions: Primary culture of rabbit fibroblast-like synoviocytes by improved explant culture method were successfully established. The method was simple and high efficient. It provided a new method for the isolation of FLS.