Hi,
I am interested in examining the effects of an endotoxin on microglial cells. I will be harvesting brains from mice at p2, treating them with this factor and then performing some molecular assays (qPCR, ELISAs, Western Blot). At the moment, a hindering factor is that we must also collect tissue for mice genotyping at the same time we collect brains for cell culture (i.e. p2) and figure a way to temporarily store brains for approximately 2 days until results are back.
I'm new to primary glial culturing & my concerns are :
1) has anyone done this and would you kindly share your method of storing?
And 2) Do you have any insight if this would affect the integrity of the tissue, stress the cells, or in any way affect microglia yield?
Thank you.
Hi Rania, you could do these things at the same time.
It takes more than 2 days to extract brain cells and get them to grow.
Hi Steingrimur,
Thanks for your feedback. For my project, we prefer not to begin growing cells until we get our mice genotyping results back so we can grow cells from selected brains based on the genotyping results. The issue is, if don't do so and randomly pick pups' brains ahead of getting our genotyping results, then there's a chance we may not have both genotypes we are testing for and so we would be wasteful with our resources...
Hi Rania, I don't know a way to keep cells happy and viable in tissue sections for days.
But growing out cells from brain sections can be done in 24 or 48 well plates.
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