Hi Everyone. I am trying to check the neuroprotective activity of some compounds in the rotenone model of Parkinson's disease in PC-12, for which I will check the effect of that compound on cell viability first. I want to ask whether it is required to differentiate PC-12 for this purpose, as I tried to differentiate the cells using 30 ng/mL as well as 50 ng/mL NGF. I am able to see the differentiated cells in 3-4 days in 96 well plate, but NGF itself is causing cell death, which could be a variable for the cell viability test.
Hi,
PC12 cells in the undifferentiated state are not neuron like. Since you want to check the neuroprotective effect of compounds the cells must be differentiated into neuron like cells, only then should neuroprotective assays be performed.
You mentioned differentiating with NGF is causing death,this is however quite unlikely.
NGF is a highly supportive differentiating agent and leads to beautiful neuronal processes in PC12 cells.
I think you must check the number of cells that you plate for differentiation. if there are too many cells they will definitely die.
if the intention is to differentiate cells, then there must be enough space in the well for the processes to grow. Hence while plating, you must seed lesser cells as compared to what you usually follow (to achieve confluency).
Cells thus plated, with NGF should differentiate well and not die.
All the best !
Hii Suriaya. I am currently seeding 5000 cells per well in 96 well plate. I will try with 2000 cells per well for this time. Moreover, I am also getting cell death in 6 well plate, where I am seeding 5,00,000 cells per well. Please suggest if I can differentiate using less dose of NGF.
Thank you
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