FRTL Cell Growth Medium
FRTL Cell Growth medium is based on Ham’s F12K medium and supplemented with hormones, L-Glutamine and FBS. Together with the TSH stock solution coming with the medium, it is ideally suited for propagating FRTL and FRTL-5 cells. Directly before usage, add the respective amount of TSH. Each aliquot provides enough TSH for preparing 100 ml complete FRTL Cell Growth Medium with a final TSH concentration of 16.5 ng/ml.
Cell lines cultured using FRTL Cell Growth Medium
Phase contrast images of FRTL-5 cells: FRTL-5 cells growing in FRTL Cell Growth Medium without TSH (right) and supplemented with 16.5 ng/mL TSH (left) 8 days after seeding.
- The FRTL Growth Medium is a basic medium, which must be finalized by adding the supplement mix provided in frozen 15 mL tubes.
- It is recommended to prepare 100 mL of medium. Take out 90 mL basic medium and add 10 mL of supplements from one of the 5 tubes, defrosted at 37°C for a maximum of 5-10 minutes. Although the supplement mix is a sterile solution, we recommend filtering it through a sterile 0.2 µm pore PES membrane syringe filter. The volume of 100 mL is sufficient to start culturing the cells and for the first subculture steps.
- Adding 10 mL supplement mix to 90 mL basic medium will result in a TSH final concentration of 16.5 ng/mL.
The proliferation of FRTL-5 cells depends on the presence of TSH in the cell culture medium
TSH-dependent FRTL-5 cell numbers. FRTL-5 were seeded at the same cell density in a medium without TSH (blue bars), containing increasing TSH concentrations (orange bars), and harvested after 8 days of incubation at 37°C/5% CO2. Cells were counted using the DeNovix Celldrop electronic cell counter in the presence of Trypan Blue.
- Basic Medium: Keep refrigerated at +2°C to +8°C in the dark. Freezing and warming up to +37° C minimize the quality of the product. Do not heat the medium to more than 37°C or use uncontrollable sources of heat (e.g., microwave appliances).
- Supplements: Keep frozen at -20°C in the dark.
- pH = 7.2 +/- 0.02 at 20-25°C.
- Each lot has been tested for sterility and the absence of mycoplasma and bacteria.