四种不同处理方法体外培养大鼠滑膜细胞*☆

背景：常用的原代滑膜细胞培养方法主要是组织块培养法、酶消化法、机械分散法等。 目的：用4种不同方法处理大鼠滑膜组织观察体外培养中滑膜细胞生长情况，探索大鼠膝关节滑膜细胞体外培养的最优方法。 方法：将大鼠滑膜细胞组织块组与剪碎组分别按4种不同的方法处理：①不加消化酶消化。②加Ⅱ型胶原酶消化。③加胰蛋白酶消化。④先加Ⅱ型胶原酶消化、再加胰蛋白酶消化。各组处理后分别进行自然沉壁和人工贴壁培养，利用倒置显微镜观察不同方法下滑膜细胞的生长状况。 结果与结论：培养9 d后，组织剪碎不加消化酶人工贴壁培养的细胞得数为2.03×106，是所有培养方法中细胞的数最多的，细胞活力为95%，细胞活力最强，优于自然贴壁。提示组织的形状、所加入酶的成分以及沉壁的方式均对细胞的生长有重要影响，大鼠膝关节滑膜细胞体外培养的最优方法为组织剪碎不加消化酶人工贴壁培养。

In vitro culture of rat synovial cells with four different treatment methods

BACKGROUND: The common method to culture the primary synovial cells is tissue cultivation, enzyme digestion and mechanical dispersion method. OBJECTIVE: With four different methods to handle big rat synovial organization in vitro, and to observe the growth situations of synovial cells and explore the optimal method of knee joint synovial cells culture in vitro. METHODS: The organize block groups and the shear ground groups were treated with four different approaches: ①Digested without enzyme digestion; ②Digested by using collagenase type Ⅱ; ③Digested by using trypsin; ④Digested sequentially by using collagenase and trypsin. After treatment, groups were cultured according to natural link wall and artificially stick wall modes, respectively. Growth situation of synovial cells were observed by using inverted microscope. RESUTLS AND CONCLUSION: Nine days later, the number of cells in organization shear group treated without enzyme digestion and artificially stick wall was 2.03×106, and that was the largest number in all the culture method. The survival rate was the highest which could reach to 95%, which was better than natural link wall. The shape of the tissue, concentration of the enzyme and the way to link played an important role on cells culture. The optimal method to in vitro culture rats knee synovial cells was tissue shear ground without enzymes dealing and cultivated with artificially stick wall.