I received SH-SY5Y WT cells and started growing them in low glucose DMEM with FBS and S/P. After reading article about changed cell morphology and detachment of cells from growth surface due to low glucose DMEM treatment I am thinking of switching to high-glucose DMEM. I have just read atcc guidelines which mention MEM as a suggested media. What are your recommendations? What do you use for your SH-SY5Y to keep them happy?
I have also frozen few vials of SH-SY5Y although I used 90% FBS and 10%DMSO instead of complete growth medium supplemented with 5% (v/v) DMSO, as atcc suggest. Should my cells be fine or would it be better for me to repeat the procedure?
I will be very grateful for all tips, I'm just starting my PhD project :)
Hey Dominika,
i have, unfortunately, no experince with your cell type, but we use for our ex vivo peritoneal macrophages (which are very delicate and littel diva's) for culture conditions in wells high glucose DMEM + 10% heat-inactivated FCS/FBS.
For four to five days, they are quite happy, then they start dying.
When we freeze cells of any kind (Macrophages, HeLa, MEFs) we use FCS/FBS with 10% DMSO (first -80°C overnight, then transfer to -150°C).
We thaw them by adding 1 ml warm medium (the optimal medium for each cell type of course) and then transfering the warmed up cells in a bigger volume to dilute the DMSO (e.g. 1 ml cell sample in FCS/DMSO + 1 ml warm medium in 8 ml warm medium). Of course, you will always have losses, but you get nearly all your cells back alive for experiments.
I hope that helped you :)
All the best,
Marc
Dear Marc Herb and Zoë P. Van Acker ,
Thank you very much for your replies! I think I will switch to high glucose medium then. I fully agree with Zoe's suggestion to freeze cells on different dates- it's always smart to have a back-up!
Best regards,
Dominika
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