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Primary Dermal Fibroblast isolation - (Jul/16/2012 )

Hi guys,

Lately when I isolate mice DF, the yield of cells is low and cells won't proliferate. The fibroblasts become large and striated (senescence).

I use this protocol:

However at the end I do an extra spin step to remove the collagenase.

Do you guys have any solution or ideas?



However right now I am not sure whether these cells are going senescent since these cells are only Passage 0-1. So I am thinking that these cells might be differentiating to myofibroblasts...

Anyone has suggestions on this or how to prevent this?