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How to grow primary neurons
Posted by: Carmen Chan (IP Hidden, Unregistered user, )
Date: March 15, 2005 07:43PM
Hi!
I'm trying to do some astrocyte-neuron co-culture. I think I've got the first part, growing astrocyte monolayer, down. But could someone please share with me a working protocol of how to grow dissociated primary neurons on the astrocyte monolayer? I was told that it's easier to grow DRG neurons compared to corticol neurons. Is that true? Perhaps I should try growing DRG neurons first? Should I use papain instead of trypsin? Should I keep cells on ice whenever possible or room temperature after trysinizing? Carmen
Re: How to grow primary neurons
Posted by: biobook (IP Hidden, Unregistered user, )
Date: June 23, 2005 12:44PM
Neuron are difficult cell type to culture. You may want to check out one of following books discussing neura cell culture.
[biowww.net]
Re: How to grow primary neurons
Posted by: sia (IP Hidden, Unregistered user, )
Date: June 28, 2005 05:28PM
Trypsin-EDTA contains other enzymes too such as chymotrypsin etc therefore you should use pure trypsin. Be gentle with the cells. Centrifuge at low speed, use fire-polished pipettes.
Re: How to grow primary neurons
Posted by: sia (IP Hidden, Unregistered user, )
Date: June 28, 2005 05:32PM
Out of those five books which one is best.
Re: How to grow primary neurons
Posted by: Glenn (IP Hidden, Unregistered user, )
Date: June 28, 2005 07:01PM
We used to order customer made trypsin from commercial vendors. Be gentle when trypsinize neuron/glial cells. Never leave them too long in trpsin.
Re: How to grow primary neurons
Posted by: h_e2000 (IP Hidden, New member, 2)
Date: August 17, 2005 01:08AM
I want to know the reason let coverslip of neuron inverted to face astrocyte in Banker co-culture system in hippocampal neuron culture. It's advantage is to let astrocyte to fall down and pure neuron? The difference between inverted coverslip and un-inverted? Thank you! Hope receive help?
h_e2000
Re: How to grow primary neurons
Posted by: hsp (IP Hidden, New member, 1)
Date: May 22, 2007 12:38AM
Hi all
I am completely new to cell culture and would really appreciate any advice offered. We would like to grow neurons and use them for subsequent cell fractionation procedures. As the phenomenon we are trying to observe is dependent on the stage of development, it would be ideal if we could get the neurons to be at approximately the same development stage in each well or flask. Does anyone know if this is possible, and what would be the best way to go about it?
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