Raw 264.7 cells- die after splitting. - (Jun/14/2007 )
Dear all,
I have a big problem with my Raw cells.
I am culturing them in RPMI+10% FCS+P/S.
And when I split them I use scrapers. I keep them in dishes and if I want to split , lets say 1:3
I suck off the medium, put 6ml new medium,. Scrape it in this medium and then take 2ml of 6ml cells and then put it in another dish which has been added 8ml fresh medium.
They have a problem in attaching and they tend to clump.
I dont know what to do.. I am really desperate. I would be grateful for any suggestions.
BEst
-clementine-
QUOTE (clementine @ Jun 14 2007, 01:35 AM)
Dear all,
I have a big problem with my Raw cells.
I am culturing them in RPMI+10% FCS+P/S.
And when I split them I use scrapers. I keep them in dishes and if I want to split , lets say 1:3
I suck off the medium, put 6ml new medium,. Scrape it in this medium and then take 2ml of 6ml cells and then put it in another dish which has been added 8ml fresh medium.
They have a problem in attaching and they tend to clump.
I dont know what to do.. I am really desperate. I would be grateful for any suggestions.
BEst
I have a big problem with my Raw cells.
I am culturing them in RPMI+10% FCS+P/S.
And when I split them I use scrapers. I keep them in dishes and if I want to split , lets say 1:3
I suck off the medium, put 6ml new medium,. Scrape it in this medium and then take 2ml of 6ml cells and then put it in another dish which has been added 8ml fresh medium.
They have a problem in attaching and they tend to clump.
I dont know what to do.. I am really desperate. I would be grateful for any suggestions.
BEst
Dear Clementine,
If I had a pound for everytime this subject comes up then I would be a rich man. RAW 264.7 SHOULD BE GROWN AS A SUSPENSION CULTURE. These are murine macrophages which stick like SUPERGLUE to Tissue Culture treated plastic. SCRAPING is no way to subculture any cell line. Trypsin will not work either.
RAW cells are almost impossible to kill if you grow them in suspension. They grow like weeds therefore large numbers of cells can be obtained every day for experiments. USE STIRRER BOTTLES and PLATFORMS i.e. Techne.
-Rhombus-