I am entirely new to cell culture studies and I need clarification on determination of seeding number after cell count with Neubauer hemocytometer. Say I have a cell density of about 4x10^6 and I need to Passage cells in a 60mm dish, what determines the seeding number per dish. I read somewhere that seeding number for a 60mm dish is 0.8 x 10^6. Is this a constant value or a discretionary value?
Hello Ayodeji Abijo
You will have to decide on the seeding density in such a way that the number of cells you decide will help to produce healthy attachment and growth. This number is unique to each cell line, meaning different cell types will have different seeding density requirements based upon their growth characteristics.
Plating cells with different densities can influence the phenotypic characteristics and the growth rate of the cells themselves. If cells are seeded at too low a density, they will go through a lag phase of growth, grow very slowly, or die out completely. If cell densities are allowed to become too high, the cells may exhaust the nutrients in the medium and die abruptly.
The seeding number of 0.8 x 10^6 which you have read for 60mm dish will vary for different cell types depending on their growth characteristics.
Best.
Hello Malcolm Nobre , Thank you for your response, I understand better now. The cell line is HEK293T. My colleague has recommended seeding density of 2x10^6.
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