Hello all,
I have been working with the J774A.1 cell line for over a year now and have succesfully thawed them from Liquid Nitrogen several times. But the past few times they are not adhering at all to the dish even though there are a lot of cells in the flask. The few that do (1-5%), I change the media and they remain attatched but don't seem to grow or multiply at all.
Do you have any suggetions? I tried using 20% in DMEM to boost them at the beginning and collagen coating the plate to try and promote rapid adhesion.
Do you have any suggestions? Thanks for any help! I'm getting a little desperate!
Best,
Anne
It may have been a bad freeze...
Laminin coating may also be an alternative.
Best wishes,
Charles
Jordan R. Yaron : We usually do not coat the plate, but I was a little desperate. I usually take them directly from liquid nitrogen and place them in the 37°C water bath. I then dilute them in 9mL 10%DMEM once the cells have started to defrost but are not 100% liquid. Then I re-suspend them and plate them straight away, without spinning them.
Charles D Anderson : Thanks!!
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