Hello S M Abdus Salam

You may use the technique of immunomagnetic cell separation. The cancer cells/fibroblasts cell suspension can be magnetically labelled with anti-fibroblast micro beads for a 15-minutes incubation period at 4 °C. The cells are washed, suspended in PBS and then passed through a 50 µm filter.

The cell suspension is then loaded onto MACS column, and a magnetic field is applied. The cancer cells are collected in the test tube placed under the column, and the fibroblasts are collected in the column due to the magnetic field. The magnetic field is turned off, and the bead-carrying cells (fibroblasts) can be released for recovery.

Magnetic cell sorting can take a positive or negative selection approach, depending on whether the desired cells remain attached to the walls of the column or flow through the column.

Best.

Dear @salam

You may separate the populations by sorting (FACS or MACS) for further analysis.

You can attempt using a saline solution (0.02% EDTA in PBS) to release the lightly anchored cells first. This is how researchers removed feeder layer cells from epithelial cell cultures for years. It requires one cell type to "hang on" more than the other. I would expect the fibroblasts to let go first, except cancer cells ( if you are growing carcinoma) anchor less than their normal epithelial counterpart. You should try it and see if it works. One set of cells will be in the saline, and the other set will require mild trypsin to release.

It will not be 100% effective though, because all the cells will release sooner or later. You have to find the sweet spot where most of one cell has released before the other one starts.

Good luck!