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本实验用静止培养法观察了接种细胞密度、血清浓度、琼脂或琼脂糖浓度及用于溶解琼脂或琼脂糖的不同溶剂对人脑胶质瘤细胞系SHG-44多细胞球体形成的影响。接种1～3×10~6细胞于铺有10m1 0.75～1.0%琼脂或琼脂糖-生理盐水或培养液底层的培养皿(直径为90mm)中,加入15ml含20%小牛血清的Eagle’sMEM培养液,所得到的球体大小较均匀,形态较圆,3～4天即可形成完整紧密的球体,重复性良好。本法与旋转瓶培养法相比,具有操作简便、经济省时等优点。
In this study, the effect of inoculating cell density, serum concentration, concentration of agar or agarose, and different solvents used to dissolve agar or agarose on the formation of human glioma cell line SHG-44 multicellular spheroids were observed in static culture. Inoculate 1 ~ 3 × 10 ~ 6 cells into Petri dishes (diameter: 90mm) plated with 10ml of 0.75-1.0% agar or agarose-saline or culture medium. Add 15ml Eagle’s MEM with 20% fetal bovine serum Culture medium, the resulting sphere size is more uniform, round shape, 3 to 4 days to form a complete tight sphere, good reproducibility. Compared with the rotating bottle culture method, this method has the advantages of simple operation, economical and time-saving.