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Doubt: Cell plating for different cell culture plate formats - (May/26/2013 )


I would like to plate cells to larger plate formats and normally would use 96-well plate (growth surface area ratio) as baseline for scaling up the plating to larger plate formats (48-well, 24-well, 12-well and 6-well). Let's say, I would like to consult whether I should scale up using cell no. based on growth surface ratio or cell density based on growth surface ratio?

For example, I plate 1 x 10^3 cells/100ul/well of 96-well plate, with cell density of 1 x 10^4 cell/ml. If I want to scale up to 24-well plate, then I should plate 1 x 10^3 cells x 6.0 (ratio of 24-well plate to 96-well plate) = 6 x 10^3 cells in 500ul (appropriate volume/well for 24-well plate). Therefore, the cell density would be 1.2 x 10^4 cells/ml. If using cell density for scaling up, the cell density for 24-well plate should be (1 x 10^4 cells/ml) x 6.0 = 6 x 10^4 cells/ml, meaning scaling up the cells no. and volume to 6.0 times.

So, which one is suitable? Normally, I would follow the first method. I just would like to standardize the calculation method. Do hope anyone could help.


Best regards,

-Seng Wu-

How do you define cell density? Since you said your cell density is xx cells/ml, which appears to me as cell concentration at plating. If that is the case, scaling up using growth surface area ratio is more appropriate.


Dear Pcrman,

Firstly, thank you for responding to my question. Yes, you could consider that is cell concentration (cells/ml). Scaling up using growth surface area ratio (second method as I mentioned previously)? Scale up both cell no. and cell volume?

-Seng Wu-

Scale up both cell no. and cell volume

By volume, do you mean volume of media added at seeding? You can scale up number of cells according to surface area ratio, follow general guidelines for the volume of media needed.