I grow 3T3-L1 cells for my research. In the differentiation process, I use a medium that contains troglitazone for 2 days. This has worked very well for about 1year, but the past couple of time, the cells started dying shortly after the differentiation starts. I am not positive it's from the troglitazone medium, but that's really the one big difference. Before I used IBMX.
Hi Aline,
Did you tried to switch back to IBMX? If the cells differentiate normally then something is wrong with your troglitazone-diff mix. But if IBMX-mix is not working properly either, something might be going on with your preadipocytes like mycoplasms contamination or chromosomal abberations. Try then to cultivate a lower passage clone or get a new clone from colleagues.
Hello Alexander,
I'm actually trying this right now, I should know over the weekend.
This is happening on a brand new set of cells.
I am having similar problem. But I am using IBMX. Please let me know if a lower passage clone works better.
Thanks
I am already using passage 4, so I don't think the passage is my problem.
I will try to test for mycoplasma
I tested for mycoplasma and it was negative. I finally was able to order a new vial of the cells and they are growing perfectly fine with lots of lipids.
After talking with some people in other labs on campus, the problem seems to be the freezing conditions. I was freezing and storing cells at -80C as it was easily available. but that only keeps the cells for a few months. Liquid nitrogen is the way to store cells. I know it says it on the cells' protocol, but circumstances made it that I didn't until now.
- Badges
- Science method