Greetings,
While PC-12 cells are commonly differentiated using nerve growth factor (NGF), I am going to be using PC-12 Adh (the adherent version of PC-12 cells) for a series of experiments involving NGF.
Here are my questions in case you have used NGF on PC-12 Adh cells:
- What media/supplements did you use for maintenance as well as when differentiating the cells?
- Did you find that the cells grew better in certain plates/flasks?
- What markers did you use to confirm differentiation? Neurite growth/length, gene expression, protein levels, etc.?
Thank you for your help!
Shahin
Just to avoid you a big waste of time; PC-12 Adh do not differentiate/grow neurites. I tried every possible experimental setups. They are not sensitive to NGF.
Thank you, Xue.
I stumbled across this paper. I hope you find it useful too: http://www.sciencedirect.com/science/article/pii/S089203620400039X
PC12 Cells
Growth medium: RPMI-1640 with 10% horse serum, 5% FBS, and 1% Penicillin-streptomycin
Differentiation media: RPMI-1640 with 1% horse serum, 1% FBS, and 1% Penicillin-streptomycin.
Hope it helps,
Just to avoid you a big waste of time; PC-12 Adh do not differentiate/grow neurites. I tried every possible experimental setups. They are not sensitive to NGF.
Much appreciated, Alexis.
I'm still going to play around with NGF and a couple of extra plates of PC12Adh cells I have. Will keep you posted!
Dear Shahin,
Alexis is correct. Please see publications from David D. Eveleth and Ralph Bradshaw at UC Irvine. In particular:
Nerve Growth Factor Nonresponsive Pheochromocytoma Cells: Altered Internalization Results in Signaling Dysfunction . By David D. Eveleth and Ralph A. Bradshaw. 1992. J. Cell Biol 117: 291-299.
I believe you can access the full text of this paper, as well as other publications of interest, in the citations of the publication posted here on RG:
https://www.researchgate.net/publication/20991127_Relationship_among_types_of_nerve_growth_factor_receptors_on_PC12_cells
Cheers!
Jill
Article Relationship among types of nerve growth factor receptors on...
Hi Shahin
It's better to use suspension PC 12 cells for differentiation purpose. Coat plates or flasks with Poly_L_Lysine and seed cells. Use NGF 50-100ng/ml for 8 days and you get differentiated PC12 cells . For confirmation, use mature neuronal markers such as neurofilament and Beta 3 Tubulin and check their expression .Iwould recommend RPMI1640 + 10%HS + 5% FBS +1%Antibiotic and antimycotic for growth of cells. For differentiation purpose use RPMI1640 with 1%HS and no FBS with antibiotic and mycotic.
Hope it works
Thank you all for providing such wonderfully detailed insights into this problem. I'm not working on this project anymore, but with your help, I can now definitely help those involved in it.
All the best!
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