Hi All. I need to prepare dissociated primary neuronal cultures enriched in parvalbumin (PV) positive neurons. I have experience culturing hippocampal and cortical neurons up to 21 DIV using the Banker’s protocol. However, for my current aims, I would need to prepare mouse cell cultures enriched in PV expressing neurons. It seems that most of protocols based on cortical dissociated neurons yield between 10-20% of GABAergic neurons, half of which could become PV positive, at most. Does any of you could recommend a protocol to get further enrichment in interneurons, specifically in PV positive ones? Should I prepare cultures from Striatum or MGE better than from cortex? Which brain area do you think is the best to work with as a source for this purpose?
Thank you very much in advance for all the help and advice.
For brain areas look into publications Buijs. If you are good in manipulation of small brain areas cut PV areas out. However will cost you a lot of mouse brains. PV neurons belong to the earliest to express, look into fetal ones. If they are still dividing!
Thank you so much for your kind answer Enrico! Are the first and middle name of Buijs Ruud M? About the number of brains, yes, I had thought in preparing a pool of dissociated cells from one or several litters. I agree than I should use fetal or embryonic brains. Thanks again.
Hi Anna,
as far as know, PV neurons develop postnatally, and are mostly absent from the neuronatal and embryonic cultures. You need to culture neurons from the adult mice to get the PV-enriched cultures, which is very tricky! Good luck!
Thanks a lot for your helpful answer Natalia!
What I found in bibliography on culturing PV-neurons was that embryonic mouse cortices are dissociated between embryonic days E13 and E15. Then dissociated neurons are grown cocultured, but not physically on the same surface, with previously grown astrocytes for 21-28 DIV to get mature "postnatal-like" PV neurons. As far as I know, culturing neurons from postnatal or adult mice yields much higher levels of glia, so the final culture will be a mixed glial -neuronal culture. This is one of my concerns working with tissue dissociated from adult animals :S. Thanks again!
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