Dear,

I don't know are there mechanical handling methods or not! but once I have suggested to one of my friends that he can separate transfected IPS cells in front of the untransfected fibroblast cells by the detection of the cells shape and size (they are distinguishable under microscope) through mechanically separated via careful sampling (Its really needs precise skill).

You might be able to try it once.

I hope you find it helpful.

I usually use Accutase followed by gentle mechanical trituration with a pipet tip to obtain single cells from spheroid cultures or human/mouse brain, which works perfectly fine for me. You might check some papers from the lab of Hans Clevers who are working on intestinal organoids. As far as i remember, they were also using Accutase for the maintenance of organoids. For establishing the culture, many method papers exist, where usually collagenase and/or hyaluronidase is used to obtain single cells.

Hi, I used Tryple Express combined with vigorous pipetting, after that I passed cell suspension through a 30 μm cell strainer.