小鼠和山羊子宫内膜淋巴细胞的制备

Preparation of Uterine intraepi the lial Lymphocytes in Mice and Goat

应用不同的消化方案 ,比较分析了胰蛋白酶和胶原酶对小鼠子宫及山羊子宫内膜组织的消化效果 ,结果证明 ,对小鼠子宫消化的最佳方案是 :1 2 0 IU· m L- 1 型胶原酶 37℃消化 4h,继之 4℃消化 1 0 h,收获已消化细胞后 ,对残余组织利用 2 .5 g· L- 1 胰蛋白酶热消化 5～ 1 0 min,之后迅速加入 5 %左右的 FCS;对山羊子宫内膜的最适消化条件是1 2 0 IU·m L- 1 型胶原酶 37℃消化 4h,继之 4℃消化 1 0 h;应用上述方法消化后 ,所获淋巴细胞活率≥ 95 %

The appropriate design for digestion of mouse uteri is to incubate the uterine tissue fragments with 120 IU·mL -1 collagnase at 37 ℃ for 4 h,followed by incubation at 4 ℃ for 10 h,if necessary to incubate the remaining fragments undigested completely with 2.5 g·L -1 trypsin at 37 ℃ for 5～10 min,then to add FCS(about 5%)to them immediately after incubation.For fragments of goat endometrium,only the course of 120 IU·mL -1 collagnase digestion is enough.The viability of the cells obtained via a series of courses of separation is always ≥95%,as estimated by trypan blue dye exclusion