I have a problem with detaching my cells: I use 1x Trypsin (0.5 mg/ml) with EDTA to detach HeLa cells. But after incubation for some minutes at 37 °C there are huge cell aggregates that do not seperate, even when I try to resuspend them by pipetting up and down. I tried it hard, but no chance. Moreover, when I want to pellet the cells, the aggregates are not soluble and because of that, I'm not able to centrifuge them down. Besides that, the suspension of trypsin and cells shows a strange behaviour on the plate: when I try to take out the suspension from the plate with the pipet, a big amount of suspension tries to redistribute on the plate even if I hold the plate slantwise. Perhaps some strange effects of surface tension.
I hope somebody can help me. Perhaps its a simple mistake I am doing, because I am relatively new in that cell culture things. My colleagues say a little bit of that 'redistribution effect' is normal.
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