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<< EYE love U护眼精灵程序下载MTT试验——细节决定成败 >>

材料:


新鲜脐带

方法:

冰冷无菌D-Hanks液中漂洗,用眼科剪仔细清除脂肪、包膜、血管等组织,转入青霉素小瓶中,加入少量无菌D-Hanks液,用眼科剪剪成0.1-1.0mm3大小的碎片,用滴管轻轻吸出上层细小脂肪碎块和油滴,再用无菌D-Hanks液反复清洗8~10次,加入10倍体积无菌胶原酶消化液(1mg/mL),室温消化30min,1000rpm离心5min,弃上清,加1mg/mL胰蛋白酶37℃消化20min,1000rpm离心5min,弃上清,再加入胰蛋白酶37℃消化20min,1000rpm离心5min,弃上清,加胶原酶(1mg/mL)弹性蛋白酶(0.5mg/mL)混合消化液37℃消化20min,1000rpm离心5min,弃消化液,再加混合消化液37℃消化20min,1000rpm离心5min,细胞加10%FBSDMEM培养基重悬,转入培养皿37℃5%CO2培养箱中培养即可。

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