some questions about hiv-1 - (Jan/31/2008 )
can anybody please explain how I can precipitate hiv-1 in cell culture medium by using peg (which rpm, how long etc.)
another question is about C8166 cells. I recently received a vial of C8166 cells from centre for aids reagents, UK. I am culturing the
cells in RPMI medium with 10% FBS but in the next morning I realize that all the cells die. this happened 3 times and the cell stock is
melting. no contamination and no specific mistakes as far as I know. can you suggest anything about this?
for rapid screening of anti-HIV-1 compounds, I'm planning to check MTT assay on CEM.SS cell line. according to a paper, this cell line
is suitable for this experiment. does anyone have any experience about this cell line?
finally can anyone send me a photo of syncytium formation? because I can see some giant cells without infecting with HIV, so I am not
sure how to detect the syncytium formation.
I know I asked too many questions but I am in trouble with these.
thanks in advance
C8166 are pretty easy to culture in standard RPMI, sure your thawing technique is good (removal of DMSO)? What experience do you have with other cell lines? They are very comparable to MT-4 (also very often used for HIV-work).
Newer worked with CEM.SS but with CEM and CEMx174 and these are easy to culture in plain RPMI and regular subculturing. Just folow instructions of how others have used these cells.
I don't have pictures of syncitium formation, but you should know that this phenomenon is and how it looks is cell line dependent, as well as virus-dependent (mostly the envelope of the virus).
In CEM-cells you should look for big "HALO-like" extensions of cells, that reach the size of severall times a normal cell. It's kind of hard to tell, but you will know by looking at it when you have syncitia and CEM are among the easiest ones to find syncitia in when compared to other cells.
thank you very much for your help vairus.
I applied normal procedures for C8166 as I did for other suspension cells but there is something wrong with them that I can't figure out.
anyway, I guess I have to culture once more and see what happens.