neuronal cultures

[Guest]

Hi,

does anyone work with hippocampal or cortical adult neurons? What are the
main problems vs embrional or postnatal cultures? It is difficult to have a
neuronal preparation not contaminated by microglia?

Thank you everyone.

[Guest]

Adult cortical neuronal cell culture? Yes, it is difficult to get any
live neurons in your cell culture. We always work with either E16-17
or P1-3. For adult neuronal culture you need to do all kinds of
dissociation, gradient separation, and neurotrophic stuff (at least in
my experience). Do some searches for adult neuronal culture methods
and I think you'll see what I mean.

[Guest]

we had also severe problems with the neurons from rat embryos.
This changed a lot, when we were testing Neurobasal A medium from Invitrogen (please remember, I do not intend to do a PR show here, it's only help!!!).
We tried many things, like low oxygen, low osmolarity, sandwich coverslip culture, astrocyte feeding layer (which seems to be really great, but we have problems with the layer...), different media from many companies and suffered through all this, sometimes great cultures, sometimes only dead cells in the well / dish.
After all we started using this literature-source as background:
Acute Effects of Ethanol on Kainate Receptors with Different Subunit Compositions, Valenzuela, Cardoso2, JPET March 1, 1999 vol. 288 no. 3 1199-1206
Acute Effects of Ethanol on Kainate Receptors with Different Subunit Compositions ? JPET

[loupurdie]

It would be helpful to know more about the specific question you are trying to answer. Are these primary cells, or immortalized cell lines? Do you want to purify a neuron specific protein, or just ID it? Are there any neuron-specific cell surface markers? Perhaps you could use a fluorescently labled antibody and sort the neurons from the glia using flow cytometry and then extract the protein. Or, if it is possible to culture glia separately from neurons (but just not vice versa), you could do a 2D gel elctrophoresis of the mixed poipulation and subtract the spots seen on a 2D gel of glia alone. You could also take the same approach with mass spec.

[Stephen Jones]

Hello,

I would like to say the body after embryonic development multiply by cell division to replenish dying cells and regenerate damaged tissues, adult cells is centred on their ability to divide or self-renew indefinitely, and generate all the cell types of the organ from which they originate, potentially regenerating the entire organ from a few cells. Unlike embryonic stem cells. Microglia are tissue macrophages that populate the mammalian central nervous system (CNS) very early in embryonic development. Microglial function have conclusively demonstrated their ability to acquire either neurotoxic OR neuroprotective functions.

Thanks a lot
Stephen Jones

[Stephen Jones]

Hi friends,

Now this is very good information that gets to the point, Neuronal Core facility also provides training and assistance in the real-time video-microscopy of cultured neurons. This technique is beneficial in analyzing neuronal dynamics and Axonal transport which, when perturbed, may contribute to neurodegeneration.

Thank you so much
Stephen Jones