Criteria for Subculture
Cells should be subcultured prior to confluence. The ideal method for determining the correct seedingdensity is to perform a growth curve at different seeding concentrations. This allows you to determine the minimum concentration that will give a short lag period and early entry into rapid logarithmic growth.
Medium requires periodic replacement. If the pH falls too rapidly, subculture may be required. Time since Last Subculture orRoutine subculture is best performed according to a strict schedule, so that reproducible behavior is achieved. It is essential to become familiar with the growth cell cycle for each cell line. Cells at different phases behave differently with respect to proliferation, enzyme activity, glycolysis and respiration, synthesis of specialized products, etc.
When cells require operations other than routine propagation (e.g., increasing stock, changing vessel or medium), this procedure should ideally be done at the regular subculture time. Cells should not be subcultured while still in the lag phase; cells should always be taken between the middle of the log phase and the plateau phase as determined during a previous subculture Fig. 1 (unless experimental requirements dictate different timing)
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