Why cultured BMDCs attach to the T75 Flask?

10 September 2020 1 5K Report

Hi,

I used GM-CSF to culture BMDCs but got some problems. On day 1, I seeded the cells in 10mm plate (106/ml) in R10+10ng/ml murine GM-CSF. Medium was changed on day 3 and day 5. On day 6, unattached cells were collected (At this time, the attached cells were about 50% confluence). I run a flow to check the quality and the results show 95% cells were live and 85% cells were CD11bCD11c double positive. Then I resuspend the cells in RPMI1640 (without FBS) to starve the cells for 2h. But even after 1h almost all of the cell attach to the bottom of the T75 flask. Anyone has seen this befor and why?

Thanks

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