HepG2 cells are a human hepatocellular carcinoma cell line widely used in various research applications. To maintain a healthy HepG2 cell culture, it is essential to use the appropriate complete growth medium. A typical complete medium for HepG2 cells consists of the following components:
- Basal medium: The most commonly used basal medium for HepG2 cells is Dulbecco’s Modified Eagle Medium (DMEM), which provides essential nutrients, vitamins, and salts to support cell growth.
- Fetal bovine serum (FBS): FBS is added to the basal medium to provide additional nutrients, growth factors, and hormones. Typically, 10% FBS is added to the medium, but the concentration may vary depending on the specific needs of your cell line and experimental setup.
- Antibiotics: To prevent bacterial contamination, antibiotics such as penicillin and streptomycin can be added to the medium. A common concentration is 100 units/mL penicillin and 100 μg/mL streptomycin.
- Additional supplements (if needed): Some protocols may require additional supplements, such as non-essential amino acids, L-glutamine, or specific growth factors, depending on the experimental conditions or objectives.
To prepare the complete medium for HepG2 cells:
- Start with the appropriate volume of DMEM.
- Add 10% FBS (e.g., 50 mL of FBS for 500 mL of DMEM).
- Add antibiotics if desired (e.g., 5 mL of 10,000 units/mL penicillin and 10 mg/mL streptomycin stock solution for 500 mL of DMEM).
- Add any additional supplements as needed.
- Mix the medium gently and sterile filter it using a 0.22 μm filter.
- Store the complete medium at 4°C for short-term use (up to 2 weeks) or aliquot and freeze at -20°C for long-term storage.
Before using the medium, equilibrate it to room temperature or warm it to 37°C in a water bath. Make sure to check the medium for any signs of contamination or discoloration before using it for cell culture. Always practice aseptic techniques to maintain the sterility of your cell cultures.
