I am trying to order media for proliferation and differentiation of PC12. I know the medium without HEPES will work just fine, but HEPES would be beneficial for maintaining pH. I don't suspect that the buffer would significantly alter anything, but I figure I'd ask before I waste money. So my question is: has anyone used RPMI 1640 GlutaMAX with HEPES and obtained good results with PC12?
Hi Kyle,
I would be surprised if it changed anything.
Please find attached an article, which uses DMEM/F12 with 0.5 % HEPES. Their differentiation seems to be normal, so i quess you can use it.
I am using regular RPMI1640 (R0883) when growing my PC12 cells and they are doing just fine.
Best regards
I have grown PC12 cell in DMEM with and without HEPES and find no difference. I like RPMI1640 better to grow the cells. I do not know whether is a personal preference or that I am still still attached to the original description to the cell line.
Good luck
Hey Kyle
I grow PC12 regularly from my experience you can go up to 1%
and still it wont make any diference.. if anything it helps preventt alowering of the ph
of the medium
good luck
Niv
Alvaro Estevez I agreed with you but HEPES is necessary and complementary component of the Cell culture Media...But depending on the type of experiment.... over all it has not any effect and also worked in a supportive manner like an additive.............
Dear Shiv I agree that HEPES can affect experimental results, which is true for any cell culture. The effects of the buffer on the results is dependent on the experimental design and the questions you are trying to answer.
About differentiating PC12 cells, in my experience RPMI, 1% horse serum and 100 ng/ml NGF work great. It takes about a week to fully differentiate, but you start seeing differentiation as soon as 3 days after the shift to the new media. This two references may help you. Greene, L. A. (1978). "Nerve growth factor prevents the death and stimulates the neuronal differentiation of clonal PC12 pheochromocytoma cells in serum-free medium." J Cell Biol 78(3): 747-755.
Greene, L. A. and A. S. Tischler (1982). "PC12 Pheochromocytoma Cultures in Neurobiological Reasearch." Advances in Cellular Neurobiology 3: 373-414.
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