RAW 264.7 cell lineculture method
Posted 08 June 2013 - 02:36 AM
I want to use RAW 264.7 cell Line. But still have some unclear problems regarding that. I will use cell line by borrowing from another lab. Those are telling me that this cell line is semi-suspension cell line, so I don't need to use Trypsin and PBS. Though when I go through web I found it is firmly attaching to the walls and need to use cell scraper.
Also I wanted to know about the serum. Some are using newborn calf serum. though i found some are using fetal bovine serum.As that FBS too expensive it would be better that if I can use newborn Calf serum
Please clarify this for me.
Thank you very much in advance.
Posted 08 June 2013 - 09:19 PM
Check the archives; I remember a discussion about 3 years ago on whether RAW’s out to be treated as attached or suspended.
Posted 01 July 2013 - 01:53 PM
Posted 01 July 2013 - 06:47 PM
However thank u very much for the replies.
Posted 18 July 2013 - 02:22 AM
I worked with RAW 264.7 cells in the late 1980's as a source of inducible nitric oxide synthase (iNOS). This enzyme was used to screen potential iNOS inhibitor compounds made synthesised by our chemists. My initial take was to optimise the induction conditions in order to produce the most iNOS for the screen. This involved:
Choosing the lipolysaccharide to use: Salmonella typhimurium (10ug/ml)
Choosing the dose of interferon gamma to use (50 units/ml)
Choosing the optimised stirring speed (cells were grown and induced in suspension culture)
However one of the most important optimisation factors was thge SERUM. We used exclusively New Zealand origin FBS/FCS as in our hands this serum gave us 50-70% more induction of iNOS than using other origin FBS/FCS.
The RAW cells are an adherent cell line and stick to tissue culture plastic like superglue. Thus we grew our cells in Techne Stirrer bottles that are made out of borosilicate glass.....this means that the cells DO NOT stick to the glass.
Hope some of this is useful