10 Useful Numbers to Remember When Starting a Cell Culture
Starting a cell culture can be an exciting but also challenging task that requires proper planning and attention to detail. Here are ten useful numbers to remember when starting a cell culture that can help increase your chances of success.
1. Cell Density
The number of cells you seed into your culture dish can greatly affect cell growth and viability. A common starting point is to seed 1×10^4 to 2×10^4 cells/cm², but this can vary depending on the cell type and culture conditions.
2. Media Volume
The optimal media volume for your culture dish also depends on cell type and culture conditions. A general rule of thumb is to use 5-10 mL of media per 25 cm² culture dish.
3. Incubation Temperature
Most mammalian cell cultures require a temperature of 37°C. However, some cells may require different temperatures, such as 33°C for some insect cells.
4. CO2 Concentration
Cell cultures require a specific CO2 concentration to maintain pH and prevent acidosis. For most mammalian cells, the optimal CO2 concentration is around 5%.
5. Passaging Frequency
Passaging refers to the process of transferring cells from one culture dish to another to prevent overgrowth and maintain cell viability. The frequency of passaging depends on the cell type and growth rate, but a general guideline is to passage cells every 3-4 days.
6. FBS Concentration
Fetal bovine serum (FBS) is commonly used as a supplement in cell culture media to promote cell growth. The optimal FBS concentration can vary depending on the cell type, but a common range is 5-10%.
7. Antibiotic Concentration
Antibiotics are often added to cell culture media to prevent contamination. The optimal concentration depends on the antibiotic used and the type of cells being cultured. A common concentration is 100 µg/mL of penicillin-streptomycin.
8. Passage Number
The passage number refers to the number of times cells have been transferred from one culture dish to another. As cells are passaged, they may accumulate genetic abnormalities or undergo senescence, so it’s important to monitor the passage number and determine when to start with fresh cells.
9. Seeding Density for Different Cell Types
Different cell types may require different seeding densities to grow optimally. For example, some cell types may require higher seeding densities than others to reach confluency.
10. Doubling Time
Doubling time refers to the time it takes for cells to double in number. This can vary depending on the cell type and culture conditions. Knowing the doubling time can help you plan when to passage cells or when to harvest cells for downstream applications.
In conclusion, starting a cell culture requires careful consideration of various factors. By keeping these ten useful numbers in mind, you can increase your chances of a successful cell culture and obtain high-quality data for your research.
