For the immortalized endometriotic stromal cells (22b), we have maintained them on an array of media types, such as Dulbecco's Modified Eagle Medium. Nutrient Mixture F-12 with 10% fetal calf serum and antibiotic/antimycotic solution and Rosenwell Park Memorial Institute 1640 medium with 10% fetal calf serum and antibiotic/antimycotic solution. We have even tried the same media with 10% fetal bovine serum as well. Unfortunately, our 22b cell line grows so slowly that it takes months to almost achieve a monolayer.
Any recommendations to help grow and maintain these cells?
The DMEM in 5%CO2 has been used successfully elsewhere.
You could try 15%FCS and do not seed your wells too thinly as that slows proliferation. Transfer some of the medium that has had cells growing in it in the hope that it contains growth factors that will speed up proliferation and try culturing some of the cells without antibiotic/antimycotic solution.
it might also be worth checking for mycoplasma infection in spite of the precautions you are taking. Mycoplasma does slow down the growth rate of cells.
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