Prinamry rat neuronal culture - (Jun/27/2005 )
I am trying to grow primary culture from adult rat brainstem (Nucleus ambuguss). Does any one know a good protocol. I know nasal and olfactory primary cultures are easy to grow and I have the protocol for them. When I used this protocol (50/50 F12 and DMEM medium with 20% FBS) I did not see much attachment of the cells. I used fire polished pipettes. There were some attchment of cells when I did not use trypsin but mostly they were not neurons (by appearance). If any one has done this type of primary culture with success in their hand please write some tricks.
-sia-
If some one has done other primary culture from other part of adult brain (other than olfactory) that would also help me.
-sia-
QUOTE (sia @ Jun 28 2005, 06:13 AM)
I am trying to grow primary culture from adult rat brainstem (Nucleus ambuguss). Does any one know a good protocol. I know nasal and olfactory primary cultures are easy to grow and I have the protocol for them. When I used this protocol (50/50 F12 and DMEM medium with 20% FBS) I did not see much attachment of the cells. I used fire polished pipettes. There were some attchment of cells when I did not use trypsin but mostly they were not neurons (by appearance). If any one has done this type of primary culture with success in their hand please write some tricks.
Dear Sia,
I also doing same thing but til now cannot establish the cell. we could discuss our method and media used.
-Jeo-
QUOTE (sia @ Jun 28 2005, 11:00 AM)
If some one has done other primary culture from other part of adult brain (other than olfactory) that would also help me.
what do u need to know for neuronal cell culture ? I have done primary hippocampal cultures.
We use neurobasal media with supplements (B27) for neuronal cultures. we c that using FBS promotes growth of non-neuronal cells so we avoid FBS completely.
-scolix-