New to Cell Culture and need some basic information - help needed (Apr/15/2008 )
How much cells do you put in for 6 wells plates for bacteria infection?
why not try a simple growth curve experiment to know your cell better?
plate some cells (minimal) in 6-well, trypsinised everyday and do a cell count.
If you get your cells fr ATCC or other culture collection, they should give a description about the doubling time. See if what you observe same with the description. If no, maybe the cells under stress or not healthy. maybe you should consider get a new stock.
In this way, you can also estimate how many cell to reach confluence in a 6-well.
Also, I've read in QIAgen RNeasy manual, there is info regarding how many cell is recommended to seed in different well plate. you can try to check there.
do you think of infection with Hp? I know these kind of experiments to infect gastric adenocarcinoma cell lines but I have difficulties to understand the usefulness; I do not believe that gastric (adeno-)carcinoma cell lines are the appropriate cell model;
as neither AGS nor MKN-74 form epithelia (they build no barrier), therefore, in my point of view, it does not matter if the cells to infect are confluent or not. If you like to compare your data to existing reports, follow their methods.
I would start with 200.000 cells per well of a 6 well plate