Question

-1. Look up the recommended centrifugation settings for HepG2 cells (time and rpm?).

-2 Calculate how much of your original cell solution you need to diluted to the volume found in question 11 to plate at the right density.

Answer 1

While cell culture is performed for established and attached cell lines such as HepG2, the process for their multiplication involves digestion using trypsin-EDTA for 5-10 minutes followed by their washing using complete media i.e. media containing basal growth medium with serum. This washing with complete media invovles brief washing of the culture plate with complete medium. This is because the action of trypsin enzyme is halted only when complete medium is added into it. The growth factors and ions present in the serum block the activity of enzyme trypsin and thus prevent any deterioration of the cell in the culture plate by it.

Hence, it is mandatory to wash the cells with complete medium while detaching.