I will start to cultivate RAW 264.7 macrophages in our lab. I have never done this before so I basically don't know anything. I have prepared a medium which is DMEM with 10% FCS and 1% gentamycin. Some have HEPES but my boss told me its not necessary. The DMEM bottle we have in our lab contains L-glutamine already, does this matter for the cultivation of RAW 264.7?
Secondly, how often will I need to split them if I thaw them and keep them in a T75 flask? How fast do they grow? Is there anything specific I should think about when thawing them?
I grow RAW264 macrophages in D-MEM supplemented with 10%FBS, 50 IU/ml of penicillin, 50 µg/ml of streptomycin, 1% sodium pyruvate and 1% non-essential amino acids. T-75 works great and for good performance you can only use these cells up to three or four passages. Beyond that these cells starts showing inconsistent results. Therefore, keep a lot of early passage RAW264 cells frozen stocks in liquid nitrogen. No need for gentamycin or antifungal drugs.
- Badges
- Science topic
- Similar topics
- Phytochemistry
- Extracts