小中大
4. 水化基底膜
用无血清培养基轻洗凝胶 (Gently wash gelled matrigel with warmed serum free-culture media.)
5. 准备细胞悬液和小室
(1). 消化法从细胞培养瓶中获取细胞; (Harvest cells from tissue culture flasks by Trypsin/EDTA;)
(2). 用培养基洗3遍(Wash the cells 3 times with culture media (RPMI1640, EMEM, DMEM etc) containing 1 % FBS.)
(3). 重选细胞,5×105 cells/ml,1% FBS (Resuspend the cells in media containing 1% FBS at a density of 5×105 cells/ml.)
(4). 上室加200 ul细胞悬液 (Put 200 ul of the cell suspension onto the matrigel.)
(5). 下室中加入600 ul细胞培养基,含有5 ug/ml fibronectin作为黏连亚族 (lower chamber of the transwell is filled with 600 ul of culture media containing, as an adhesive subtrate.)
6. 孵育,37℃,20 to 24 h (Incubate at 37C for 20 to 24 h.