Abstract: In cell culture, selection of culture bottles and the added amount of medium are direct factors for influencing cell growth. How to deal with the two factors is specified below.
Keywords: Cell Culture, Cell Morphology, Cell Growth
1. Selection of Culture Bottle
We find the cell with rapid growth rate grows better in glass bottle, comparing with disposable plastic bottle. For the same cell during its strong and rapid growth period, the comparison result of the growth state in glass and plastic bottle is still the same. Probably, the cell could attach easier in plastic bottle. However, it grows worse in plastic bottle which appears to be a more comfortable environment.
For the cell with slow growth rate, the cell morphology in plastic bottle is better than in glass bottle. Glass bottle is a good choice for the cell growing rapidly. For the same cell, plastic bottle is suggested to be used in the slow growth period. E.g. the cell just recovers or is in primary culture. Contrarily, glass bottle fits the cell growing rapidly.
2. Added Amount of Medium
The added amount of medium depends on each cell habit which should be discovered by yourself. 12ml / 14ml medium added for small or big glass square bottle is not always fixed. For some cells, the morphology can be better with less medium. (The competition may be intense.) Less medium fits the cell growing rapidly and can get a better morphology. Medium change frequency shall be mastered well.
During subculture, PBS wash is a very important step. Recently, we find cells can be dispersed by PBS. Add PBS for 10-15min or even longer for some cells. After cells are separated, they can be dealt with a pipet. Like trypsin digestion, time should be controlled well. Otherwise, cell is harmed for the overtime and the bad condition will greatly influence cell viability. This method is suitable for some cells which have difficulties in being digested into single cell by trypsin. Alternatively, the trypsin is unavailable. A trial operation is suggested for your cell.