I am using DMEM from gibco for the culturing, but cells seems not happy. I have also tried DMEM Glutamax but in all case cells do not survive more than one month. And the growth is very slow. Is anybody here has any experience in these cells? Please do me help to overcome the problem.
DMEM/F12 with 15-20% FCS should be better. I have experience only with tail fibroblasts. But may be the paper below could help you troubleshoot. They suggest reduced oxygen for better survival of cells. Good Luck.
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3185624/#!po=10.0000
Thanks a lot Manoj, I will try to apply the FCS concentration and follow the article.
did you try preparing the media with 10% FBS AND 10% pen strep as an antibiotics. then after putting the cells in the media prepared. try once per week adding aeound 1ml FBS serum .
Dear Amira, thanks a lot, i could not get it fully . should i add 1 ml FBS in 10 ml of plate. and another issue is i split them in every 3 to 4 day, so should i increase it every time.?
I mean when u get the bottle of media which is lets say 500ml you add to it 50mls fbs and then 5 mls pen strep and you shake. Thats how you prepare a good media for your plating and cell culture work
and yes you add 1 ml in each 10 mls flask.
no need to increase
Dear Amira, thanks a lot. for detailed response. i will try to follow the same.
I think that in addition of the media you use, it is important to know how are you preparing the fibroblast, are these a commercial fibroblast a cell line? or are primary cells isolated from lung tissue? Let me know!
Dear Hemant, did you have luck with the mice lung fibroblasts? I find them very difficult to expand, they tend to differentiate into myofibroblats very easily. Any tips?
Thanks
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