Hello,
I have a problem with adhesion of astrocyte C8-D1A line cells. After thawing, the cells are alive (in trypan blue staining, more than 95% viability). But they don't adhese to the bottle. Increasing FBS to 20% did not help. Iculture them on NUNC Delta-surface bottles. I would like to add that after buying from the ATCC collection, they stuck poorly, but grew. I managed to bank in liquid nitrogen and did a number of experiments. The problem appeared after I thawed the ampoules frozen by me. Does anyone have any experience with this? What could be wrong?
Thank you for your help,
Małgorzata
Dear Malgorzata!
When you freeze the cells, did you use any cryoprotectant?
For example, DMSO. Usually, the cells are frozen, 10-15% of the volume of the DMSO medium is added to the medium with serum.
Hello Malgorzata
What is the culture medium you use ? Try to use DMEM with high glucose and 10% serum. Also I suggest you use trypsin of lower concentration during sub culturing because these cells I feel are sensitive to trypsin.
Thank you for answers! :)
Aleksandr, yes I use 10% DMSO to frozen medium.
Malcolm, I culture my cells in ATCC-formulated DMEM, according to ATCC recomendation. Trypsin, is, according to recomendation too, is 0,25%. But I don't get to that stage because the cells don't adheze to the surface of the culture flask
;(
- Badges
- Science method
- Similar topics
- Biological Science
- Histology
- Staining