To subculture cells, you must plate the cells on a new plates at a specific dens

Question

To subculture cells, you must plate the cells on a new plates at a specific density that depends on that cell line and the purpose of your experiment. If you needed to plate the cells(ie. That your group resuspended in 500microliters) at a concentration of 2200 cells per cm2 onto 35mm plates that have an effective growth surface area of 9cm2, how would you do this? The final volume of media for a 35mm plate is 2mL.
Total # of 35mm culture dishes plated?
Volume of media to add to cell suspension?
Please explain throughly
Thank you

Explanation / Answer

The information regarding the initial number of cells "(ie. That your group resuspended in 500 microliters)" is misssing in the question. Hence, I will solve this question making an assumption that the total number of cells "that your group resuspended in 500 microliters" was N. So, the stock seed contains N/500 cells/L = 2N cells/mL.

Given that,

1. the cells are to be plated at a concentration of 2200 cells per cm2, and

2. the plate has an effective growth surface area of 9 cm2 (= 2 mL).

Thus total number of cells required = 2200 × 9 = 19,800 cells.

The stock contains 2N cells/mL. Thus, the amount of seed needed for obtaining 19,800 cells = 19800/2N = 9900/N mL

Hence, you should take 9900/N mL of seed and dilute it with [2-(9900/N)] mL of medium to get final 2 mL of cell suspension for plating the 35 mm plate.

If for example the initial number of cells "that your group resuspended in 500 microliters" was 104 (i.e. N = 104), then you must take 9900/104 = 0.99 mL (990 L) of the stock seed, dilute it with (2-0.99) = 1.01 mL (1010 L) of medium to get final 2 mL of cell suspension for plating the 35 mm plate.

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