Q: Astrocytes in culture

[S.T. Kirat]

Hi.  I'm an undergraduate working part-time in a neuroscience lab, and 
have been experiencing some difficulty with astrocytes in culture.  
Although I do not grow up the cultures myself, I am dependent on them for 
my own project, and I can't seem to get the cells in a "happy" state.  My 
attemps to research culturing techniques have been futile because most 
research publications don't give the methods in enough detail for a novice.
  As I mentioned before, my cultures are set up for me.  All I do is 
harvest the cells after the assay condition and perform Northern blots 
from the RNA I extract.  The assay condition is applied only four hours 
after trypsonization for splitting purposes, and I believe that this may 
be the problem, since after harvesting 10 x 100mm diameter culture dishes, 
the cell pellet I obtain is between 1 - 2 mm in diameter.
  Can anyone give me advice as to what is an acceptable waiting period after 
trypsonization?  Thanks in advance.


-- 
Sebnem Kirat =-)
Biochemistry IV, McMaster University
u9106353 at muss.cis.mcmaster.ca