I am trying to establish cell cultures from a sample of bladder cancer that we develop in mice but we have many fibroblasts in our cultures. Does anyone knows how to inhibit them without disturbing cancer cells?
Many thanks!
I have done this with only *some* effectiveness with differential trypsinization. Fibroblasts tend to release from the dishes at lower concentrations of trypsin than epithelial cells so if you're careful you can trypisinize only the fibroblasts and leave the other cells stuck to the plate. A more effective (but also more costly) method is to use flow or magnetic cell sorting based on fibroblast expression of CD90 (Thy) to physically remove them from a suspended cell mixture.
Thanks for your answer Jennifer! I have already tried differential trypsinization so i will keep on this. :)
Differential trypsinization will result in dislodging mesenchymal cell first. So you may miss mesenchymal cells in addition to fibroblasts. I would suggest to maintain longer so that the fibroblasts go off by replicative senescence whereas cancer cells outnumber fibroblasts over time.
You may also want to try to reduce your serum in the growth media to 1%. Depending on your cell line they will also slow down a lot but they will eventually outgrow the fibroblats.
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