I have tried different medium complements for each cell lines but there is a contamination that i don't know. So i couldn't expand them in culture. Which supplement i should use for each cell lines in medium.
Hello Elif Yaşar
For SCC-9 cell line, the growth media is DMEM:HAMS F12 (1:1) + 2mM Glutamine + 10% FBS + 0.4 ug/ml hydrocortisone + 0.5mM sodium pyruvate.
The growth of SCC-9 cells is enhanced by the use of a feeder layer of X-irradiated STO fibroblast cells.
For FaDu and Detroit 562 cell lines, the growth media is EMEM +10% FBS.
Usually, antibiotics are not added to culture. But since you are facing the issue of contamination, in addition to maintaining sterility, you may add Penicillin-Streptomycin solution at a concentration of 50-100 I.U./ml to the culture media. Antibiotics give extra layer of protection. However, after a few passages in culture, you may discontinue the use of antibiotics and strictly follow aseptic technique which is a collection of procedures and techniques designed to prevent the introduction of unwanted microorganisms into pure culture or into the laboratory environment. Maintaining sterility during culture will help prevent contamination.
Good Luck!
1. SCC-9 Cell Line:
SCC-9 cells are often cultured in Dulbecco's Modified Eagle's Medium (DMEM) or a 1:1 mixture of DMEM and Ham's F-12 medium.
2. FaDu Cell Line:
FaDu cells are commonly cultured in Eagle's Minimum Essential Medium (EMEM) or DMEM.
3. Detroit Cell Line (e.g., Detroit 562):
Detroit cells are often cultured in EMEM or DMEM, similar to FaDu cells.
To address the issue of contamination, here are some general tips:
1. Maintain sterile technique: Ensure that all equipment, reagents, and surfaces are properly sterilized to prevent contamination.
2. Monitor your incubator and water source: Regularly check and maintain your cell culture incubator, as well as the water used for media preparation, to prevent microbial contamination.
3. Use good aseptic practices: Wash your hands, wear appropriate personal protective equipment (PPE), and work in a laminar flow hood or biosafety cabinet when handling cells.
4. Regularly check for mycoplasma contamination: Mycoplasma contamination can go unnoticed but can affect cell health. Perform regular mycoplasma testing to ensure the absence of mycoplasma in your cultures.
All the best
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