I want to culture rat DRG neurons without contamination from other cells and debris from the ganglia. I tried a purification procedure using differential concentrations of Percall but the result was not satisfactory in case of cell loss and survival. I need a large number of neurons (and least trauma) in culture.
you may consult STEMCELL Technologies Inc. they have specific cell isolation kit or protocol.
Good luck.
I thank both of you very much for your answer, especially to Dr. Potter for sharing the protocol. DRG neurons I want to purify will be obtained from adult rats and the protocol I have been used is quite different. However, your suggested protocol may give me some idea.
I spin down 1 mL of the cell suspension through a column of 2 mL 15% BSA (dissolved in DMEM:F-12, but I don't think this is too important, you could probably get away with PBS) at 200 RCF for 10 minutes. This gets rid of all the myelin debris and highly enriches the neuronal population. It does not, however, get rid of ALL glia/fibroblasts.
The protocol I used (some time ago now) was obtained from Prof. Janet Keast who is now at the University of Melbourne. Best of luck!
Vipavadee,
If you are looking for DRG cells, BrainBits sells Embryonic and Postnatal DRG Cells. These come fresh, not frozen, and should be helpful for exactly what you need.
Rat Embryonic: http://www.brainbitsllc.com/embryonic-1/
Rat Postnatal: http://www.brainbitsllc.com/postnatal/
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