More questions about culturing human fibroblasts (ARGH!) - (Nov/29/2006 )
So I am still tending to these patient derived fibroblasts (yuck!). One is growing very well and looks good. A new one I received a couple of weeks ago initally grew some, I split it out of it's original flask (it was about 80% confluent at most) and then it stopped growing. Another set of cells is growing a bit better - after I froze some and took them out again, that seemed to help - but still v slowly. And finally, the last set do not grow at all, or very little as far as I can tell. My colleague informs me that someone has requested these but that will be tough to do since they don't grow! We have (allegedly) freezer stocks of these so will try to revive some and see if they are better. Some questions!
1. Is it growth media? Some were growing in BEM when I took over but I've switched all of them over to MEM + Earle's since this is what the healthy proliferating cells were growing in. I don't know if it has improved things, but it is certainly not worse!
2. Stupid question, but what do fibroblasts tend to look like 'normally'. The healthy ones have a nice even shape - sort of thin and elongaged. Some of the other cells look really icky - with large vacuoles and unknown rubbish in the cell body. The worst growing cell line is flat and square-ish with really jagged edges. What do they look like when they've reached senesence?
3. How quickly do 'normal' primary human fibroblasts grow. Or is there no normal?
Any light you could shed on this would be greatfully appreciated :-).
well, usually that is a sign of stress, and since they are growing poorly -> it is more than obvious that they must have some stress.