1. Cell culture process
Following the correct thawing procedure of frozen cells is very important for ensuring successful culturing.
- Do not allow the cell density in the dishes to become too dilute. It may restrict growth in the first culture.
- Determine viability when you open frozen vials. Especially in suspension cultures, it is difficult to separate dead cells and living cells.
- Check the mitotic rate of cell lines. Each cell line has a different mitotic rate, from about 12hr to 2 days or more.
- Follow to the instructions carefully. Differentiation and aging may cause a proliferation disorder.
- Change the culture medium at appropriate intervals. These depend on the cell lines and cell density. If the cell density is high, you should renew medium at shorter intervals. Likewise, keep long intervals if the cells are at low density.
- Subculture the cell lines at appropriate times. Post-confluent cells may undergo differenciation and exhibit reduced proliferation after passaging.
2. Guidelines for maintaining CO2 incubator
Keep the incubator clean and regularly check inside temperature, humidity and CO2 concentration. Ensure it is regularly serviced and calibrated, according to the manufacturer’s instructions.
3. Dishes and flasks
Please use appropriate cell culture dishes or flasks. Suspension culture grade or microbial grade dishes are inadequate for growing adherent cells.
The size of dishes and flasks critically affects the survival and growth of cells at the first culture. In general, each vial of JCRB cell lines is prepared to start cell culture in one T-25 flask or one 6-cm dish. Do not use dishes or flasks that are too large or small, as further growth may be adversely affected.