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Sub-culture of N2a cells - (Oct/07/2009 )

Hi all. I'm trying to grow N2a cells at the moment. I got them from ATCC and followed the instructions enclosed. However, when I attempted to sub-culture my cells I ran into problems. After centrifugation my cells were really damaged and it has taken 3 weeks to get them to a very low 30-40% confluency. I had to change from minimal media using EMEM to DMEM to the DMEM/50% Opti-mem mix I am currently using.

Can anyone tell me how to what the optimim centrifugation speed is? The reason I use this step is to try and recover the non-adherent cells and remove debris.

I really need 80-90% confluent flasks and this is beginning to depress me. :( Any help appreciated!


You shouldn't be using over 100 RCF for centrifuging