I will do mixed primary culture from neonate's brain and I for sure I will have astrocytes and neurons (with DMEM + FBS and Neurobasal + B27).
But I wanna know about the other glial cells, for how much time they could survive at the culture? For how long?
I would like the maximum heterogeneity as possible.
Does someone know?
Dear Renata,
In terms of glial cells, your cultures will also contain microglia, oligodendrocyte precursor cells, and oligodendrocytes.
One place that has some information is this review:
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2140055/
Best,
Jill
Hi Renata, what area of the brain are you using? Also what are you planning to do with those cells? I ask because you may find big differences y distinct areas and depending on the application, your culture media may need modifications. In general astrocytes may survive well in neurobasal, specially if you have serum, but neurons are not super happy with over 5% serum. I do cultures of astrocytes with DMEM, glutamax, and 10% serum from hippocampus. The cells proliferate and you get a confluent astrocytes bed in a couple of weeks (depneding on the density). Most of the neurons die. I grow neurons on top in neurobasal with b27 (no serum) and the astrocytes can handle it for at least 3 weeks. No much microglia or olygodendrocytes in my experience, but definitely there.
Thank you so much everyone!
Jill, thank you for the article! I was afraid of in a week or ten days (the time for the culture achieve 70% of confluence) the microglia and oligodendrocytes could die.
Maria, I do not wanna isolate the cells, I wanna a model the most similar to the cortex area, it why I wanna the presence of astrocytes, neurons and microglia.
Marcelo, when the cells achieve like 70% of confluence, I will put a layer of breast tumor cells (with its respective medium and serum) for two days.
Guys, do you know if in these primary cultures (from cerebral cortex of neonates) we could find some neural stem cells and for how long in vitro?
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