I would like to ask what's the best protocol for cell thawing, I noticed some people suggest changing the media after cells are attached while some suggest to change media after 24 hr. I would like to thaw some cells, does it matter if media is changed 24 hr after thawing?
Thank you.
I freeze my cells in DMEM, 20% FBS and 10% DMSO. After thawing i dilute them in 10 times more DMEM + 10% FBS to dilute DMSO. Then I pellet down cells wash them in PBS and transfer all in one tube, pellet again, resuspend in DMEM + 10% FBS+ P/S and plate them. Tomorrow I exchange their medium to get rid of dead cells and leftover DMSO.
Yes always spin down your thawed cells and resuspend with fresh culture media as Mateo suggests. I also change media the next day and have no issues whatsoever. I suppose you could change media after you have waited a couple hours after the cells have attached but in my experience (using epithelial (like)) cells, its no needed.
- Badges
- Science topic
- Similar topics
- Biological Science
- Molecular Cell Biology
- Culture Cells
- Cell Line