Other technical issues

As cells generally continue to divide in culture, they generally grow to fill the available area or volume. This can generate several issues:

1. Nutrient depletion in the growth media
2. Accumulation of apoptotic/necrotic (dead) cells
3. Cell-to-cell contact can stimulate cell cycle arrest, causing cells to stop dividing, known as contact inhibition.
4. Cell-to-cell contact can stimulate cellular differentiation.
5. Genetic and epigenetic alterations, with a natural selection of the altered cells potentially leading to overgrowth of abnormal, culture-adapted cells with decreased differentiation and increased proliferative capacity.