In ATCC website, they have an explanation hereinbelow:
(Note: The L-15 medium formulation was devised for use in a free gas exchange with atmospheric air. A CO2 and air mixture is detrimental to cells when using this medium for cultivation)
Culture Conditions: Atmosphere: air, 100%
If I use the Co2 incubator without the Co2 cylinder does it mean that I reached the desired condition? Is there anyone who worked with this cell line?
Best Regards,
Sena
To make your life easier, try to cultivate them in another medium; for suggestions see:
https://www.researchgate.net/post/Why_do_MDA-MB-231_cells_start_to_die_after_first_passage
Hello
You can use RPMI 1640 w/ 10% FBS, 2% P/S, 1.25% L-glut, and 1% sodium pyruvate or DMEM+10%FBS +NEAA for MDA-231 cells and it works great.
L-15 is designed to be used not in a CO2 controled incubator
For sub-culture : Remove medium and rinse the adherent cells using PBS without calcium and magnesium (3-5 ml PBS for T25, 5-10ml for T75 cell culture flasks). Add TrypleExpress( 1-2ml per T25, 2.5ml per T75 cell culture flask), the cell sheet must be covered completely. Incubate at 37°C for 10 minutes. Carefully resuspend the cells with medium (10 ml), centrifuge for 5 min at 300xg,
Good Luck
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