Hello,
I am trying to obtain suspension cultures of HEK293 cells in serum-free media (SFM).
I adapt them from adhesion in DMEM+FBS to SFM and transfer them to shaking flask at 120 rpm 37°C 5% CO2.
I use an automatic BioRad TC10 counter with trypan blue to count the cells and assess viability. I also look at them with a microscope.
While the total number of cells in increasing, the viability really hardly goes above 50%.
In some media, I observe some aggregates (which I suppose don't help to have a good viability).
I also have difficulties to separate dead cells from the living ones.
Any one with hands-on experience on the matter ? The majority of protocols and documentation lack detailed practical handling information.
Thanks a lot
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