I am working with U87MG (glioblastoma) cell line. I am using MEM with NEAA+ 1mM Na Pyr+ FBS as media. The cells grow fine in T-25 flasks. But when I seed them in a 96 or 12 well plate, the cells in the side wells detach with complete layer after some time while the cells in the middle wells stay attached. What could be the reason ?
I have experienced this. This is because the water of the side wells evaparate much faster than the middle wells, creating harsh enviorment for the cells to live. This is called "side effect", and is exactly why we do not use side wells to plant cells. Instead, we put culture medium (same quantity as the liquid in the middle wells) in side wells, creating moist enviorment for the middle wells. In this way, your cells will be good. Wish you success!
Hello
I agree with Liang Hong. This type of problem is caused because of 'Edge Effect'. Temperature difference across the plate and evaporation effects in the side wells during incubation may be the cause of such type of cell behaviour.
In the side wells because of evaporation salts from the culture medium could accumulate in the wells affecting cell attachment and metabolism.
I would suggest that you avoid seeding cells in the side wells.
Best.
- Badges
- Science topic
- Similar topics
- Biological Science
- Microbiology
- Virology
- Virus