Hi, I am just curious as to why I should count the cell when doing cell passage.
If I am subculturing just to maintain the cell line, I feel like there is no need to count the cell. For example, if I am to subculture a t75 flask culture to three new t75 flasks, just to maintain and keep extra cell line, I do not necessarily have to count the cell from old t75 flask, am I right?
You're likely to get answers supporting both sides of this issue, and in many cases it seems to be a personal choice amongst researchers.
I typically do not count my cells when subculturing, I split to new flasks using a generalized rule of thumb of 3 new flasks from 1 old flask. A few caveats to keep in mind though when doing it that way:
1) do not let the initial flask grow to complete (100%) confluence. Many cell lines can become growth arrested or senescent if they become overconfluent, this may have an effect on the growth of cells in the new flasks. I split mine when they are perhaps 75-85% confluent and still have mitotic figures.
2) Don't just use the "3 flasks from 1" rule blindly. Different cell lines have different growth rates, and can be seeded at different densities. When we split our B16F10 melanoma cells for example - which are very fast-growing - I'll split as high as 1 to 5. Otherwise, I'd be splitting them every other day.
3) It may be wise to count a new cell line the first few times you subculture them, and be certain to pay attention to it's growth characteristics. Some line prefer to be seeded a little more densely than others (I've had this with SKBR and LNCaP cells). Let the cell line show you what it's optimal conditions are and split accordingly from that point on.
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