I thawed a cell from the -80 freezer. I added the cell content from the cryovial onto a tissue culture plate containing 10 ml of growth media. I left the cells to incubate inside a 37 degree incubator for cell growth for one overnight. The followeing day, the media was changed. My question is, since I did not centrifuge the thawed cells to remove dmso, will this affect the cells? The media was changed only one day after the cells were thawed. the DMSO concentration is at 2%. will this affect the DNA property of the cell?
You might be ok, DMSO could be toxic to the cells and you should wash these cells always before giving them a fresh passage.
This link explains a bit more about it
http://cellculturedish.com/2011/11/dmso-cryopreservation-comes-with-a-cost/
This definitely affected your cells. Always wash unless you are diluting the DMSO out 100x.
Typically, when I removed cells from liquid nitrogen to start a culture, they are frozen in a freezing medium containing 90% FBS + 10% DMSO. Generally, cells are kept in 1 ml of this freezing mixture and once the vial has been quickly thawed, I add it to a culture flask that contains 15 ml of normal cell medium. I generally have no issues with the relatively small amount of DMSO. I work with primary human skin cells but other cells or cell lines might have different sensitivity to DMSO.
I agree with the others, it is best to remove the DMSO prior to plating. You did not mention the cell type you are using. There is a differential effect of DMSO dependent on the differentiated state of the cells. Meaning, differentiated cells are the least affected by DMSO, next are the tissue committed progenitor cells, and then there are the lineage-uncommitted stem cells, which are affected the most with DMSO. DMSO will alter phenotypic expression of stem cells if incubated with the cells at 37C, even at extremely low doses. Then there is the matter of DMSO being a universal solvent. So anything dissolved in the DMSO can gain entrance into the cell proper and alter its gene expression and subsequent physiology. And, as mentioned above, DMSO can be cytotoxic. But this cytotoxicity may not show up until the cells are passaged 1-2 times.
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