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Is there a generally accepted maximum number of times you should split and reseed mammalian culture cells before restarting cultures from new stock?

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I only know the maxime to keep the passage number as low as possible. Especially in immortalized cell lines funny things can happen, we once found a cell line in the lab (I think it was 293T) that was hexaploid for some chromosomes...So split cell lines in early passage numbers and make stocks. The problem you have with cells from other labs is that you often don't know how long the line has been in culture... –  Chris Jan 30 at 20:40
Yeah, unfortunately this was the first time I had done any mammalian tissue culturing, by the time I had grown enough to make a sufficiently large stock of frozen cells, I had already hit passage number 6. I was thinking I would draw from frozen stock each time my current line hits passage number 10. Does that seem reasonable or should I buy new cells? We sourced our current cells directly from the American Type Culture Collection. –  Dan.Riggins Feb 3 at 18:31
How long does it take to do six passages? –  Chris Feb 3 at 18:55
About 9 weeks. :/ I made a lot of dumb errors at the start. –  Dan.Riggins Feb 3 at 22:38
If your cells come from ATCC they shouldn't be in culture for ages, since they are one of the good cell banks. I would still use this cell stocks and I would also test, how long you can keep them in culture without noticing changes in behavious and/or shape. What does ATCC recommend here? –  Chris Feb 4 at 6:40

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