低糖DMEM+FBS10%培养SD大鼠MSC，P0和P1代消化不下来啊很吃力。

倒腾细胞

胰酶应该是没问题的，因为之前用于消化高糖培养基的时候很顺利。前辈们在这里指点到的胰酶放箱预热也用了，还是大部分都死贴，消不下来。  K% K( A- ?$ N
据说细胞老化的话，也很难消，下图为P1代细胞，貌似没有老化啊。还有什么原因会导致消化无力呢？望前辈们指点。
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屋檐上的乌鸦

为何要用低糖DMEM呢？

kakapo

there are lot of adipocytes and fibroblasts  in adipose tissue. When one perform digestion of adipose tissue with collagenase, the adipocytes and fibroblasts also release from the adipose tissue. Well, for MSC culture, one has to select out fibroblast and adipocytes. Metabolic way is a strategy to clean adipocytes and fibroblasts out. The adipocytes and fibroblast can not survive under condition of low glucose DMEM. Then MSC can grow in low glucose DMEM.  

zhen022

感染的死细胞？

倒腾细胞

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倒腾细胞

回复 zhen022 的帖子
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根据我的操作，细胞污染确实有可能。那是什么感染呢、

倒腾细胞

回复 屋檐上的乌鸦 的帖子
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/ B0 K" \1 H$ F: zthere are lot of adipocytes and fibroblasts  in adipose tissue. When one perform digestion of adipose tissue with collagenase, the adipocytes and fibroblasts also release from the adipose tissue. Well, for MSC culture, one has to select out fibroblast and adipocytes. Metabolic way is a strategy to clean adipocytes and fibroblasts out. The adipocytes and fibroblast can not survive under condition of low glucose DMEM. Then MSC can grow in low glucose DMEM.  

末末

我也很好奇怎么会消化不下来呢？消化之前有没有用PBS吹洗瓶壁呢？微量的FBS就能导致胰酶失效的。

frankhu

从图片上看，我不确定这些细胞就是MSCs，不管MSCs有没有老化，一般很少会发生消化不下来的情况

鹏程万里

我两周前也出现过这种情况，据我估计，这个不像是MSC，应该是上皮细胞类，我那个细胞费了很大劲消化下来，最后也全死了。