We have HUVEC bought from Invitrogen. Cat no -C-003-5c. while first plating the cells we have bought the protocol suggests not to centrifuge the cell with media to remove the DMSO. Should we remove the DMSO by centrifuging? If not should we change the media after 6 hrs ? It is written in the protocol not to disturb the media for first 24 hrs. So is it advisable to change the media after first 6 hrs?
Hello Argha
I advise you to follow the protocol. If it says not to disturb the medium for the first 24 hours, you should not because there must be a reason for this. There are protocols stating that culture medium containing DMSO is replaced after first 24 hours. How did you arrive at 6 hours ?
Good Luck!
I agree with Malcolm Nobre . I think you don't have to worry about holding cells in DMSO for 24 hours. I've done it myself, with different cell line, but the procedure is still the same.
The reason why you shouldn't centrifuge the cells is connected with the fact, that freshly thawed cells might be weaker. Centrifugation may damage them.
Also, they might need a little more time to attach to the plate. That's why we should give them a day, to avoid aspirating alive cells with medium.
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