help for Isolation of FIBROBLAST from tumor(tissue speciment) - (Sep/16/2005 )

hi all

I am in desperate need of protocol to isolate the fibroblast from tumor(tissue specimen). I need to develop cell line from tumor itself for good results. if some body can be kind enough in providing me the whole protocol, I will be much obliged.

Looking forward for positive response.


Thanks

Waraich

It is very easy to isolate fibroblasts from primary tumours. However, if you want to make a cell line from the primary tumour, you will need to isolate epithelial cells from the tumour. And that will turn out to be far more difficult since the fibroblasts will easily overgrow the epithelial cells.

if they are keratinocytes you want to isolate from fibroblasts, growing the specimen in defined KGM medium should isolate the keratinocyte epithelials from the fibroblasts (you get about 98% epithelials and there are always a few FB's, but if the epithelials are happy and not senescent they will vastly outgrow the FB's)

not sure if this works with other epithelial cell types?