Dear all,
I am starting working with E0771 cell line since I have to establish an orthotopic breast tumor model in c57 mice but I have no experience with this cell line so I would really appreciate any advice that you can give me.
In particular, I saw the ATCC website and they say that it is better to culture these cells in a t-75 corning flask, maintaining cultures at a cell concentration between 6 x 10^4 and 8 x 10^4 cells/cm2, is this true also for your experience? How many cells do you plate in a 75 cm2 flask?
Do they grow fast? How many times per week do you subculture them?
Sorry for all of these questions but I am new with these cells and so I would really be very grateful for all your advice,
Thanks a lot,
Giulia
Hi,
It doesn't matter what you culture the cells in, I culture mine in 10cm plates and they are perfectly happy. They do grow quite fast, if you use them regularly, splitting them 1:10 usually means splitting them 2-3 times a week. They are fine with being split more harshly if you don't need them as much and want to reduce the number of splits. Be careful that you don't leave them confluent for too long, as with most cells, they're not happy like that, especially since they use the nutrients in the media quite quickly.
Good luck with them!
I can classically recommend that you stock up as much as possible in the early passages. Because as the number of passages increased, I had difficulty in culturing.
Also, according to my experience; if the cells you thaw from the first refrigerator are very low confluency, you may have difficulty. Therefore, in the first step, you can start on the T-25. They seriously need cell-to-cell interaction while growing.
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