You are growing CHO cells in tissue culture and need to count them for your expe

Question

You are growing CHO cells in tissue culture and need to count them for your experiment. You dilute 2 mL cell suspension per 10 mL liquid medium and pipet a drop of this diluted cell suspension onto a hemocytometer. Looking under the microscope, you count 72 cells within the 10 grid squares.

a. What is the concentration of your cell suspension in cells/mL? Answer in decimal notation.

b. The protocol asks for a final cell concentration of 1.0e8 cells per mL. You centrifuge 250 mL of the cell suspension and remove all of the liquid supernatant from the cell pellet. In how much liquid media (in mL) do you resuspend the cell pellet to end up with a final cell concentration of 1.0e8 cells per mL? Answer in number of mL to one decimal place.

Explanation / Answer

Answer

a) The total count from 10 grids = 72 (cells/mL x1010squares )


2. Then multiply by 2 to adjust for the 1:2 dilution in trypan blue.

These two steps are equivalent to dividing the cell count by 2.

As an example:

If the cell count is 72, the cell density is:

72 = 36 x 1010/mL

b )cannot be determined as the terms are not defined the term 1.0e8 cells per mL is not clear
2

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