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Cell lysis when isolating dendritic cells - (Dec/21/2004 )

Hello everyone, I have a simple question regarding DC harvesting.

I currently use the plastic adherence technique when isolating DC precursors. So after the 2hr incubation, I am ready to pool the cells into one flask. However, it seems that if I use a cell scraper to harvest the cells, I observe cell lysis which results in big clumps of cells. Since I am working with a small amount of cells, I need everyone I can get. What is the best way to harvest the adherent cells from the flask that will result in minimal cell lysis?
These cells are isolated from peripheral blood and incubated in AIM V serum free media. Has anyone else had lysis problems when isolating DC precursors from peripheral blood?

Thanks for the help.



Instead of a scraper, do you think that incubation with trypsin for 5 min could solve your problem?

... no more than 5 min!!...