Magnetic Activated Cell Sorting (or Separation) is the best method (and to be honest, one of the few methods, I am also open to suggestions) that I know of that gives a proper yield and has a reliable purity of the cells.

Basically (in case you're not familiar): You purify your sample (because this method is usually done on full blood) and when youy add the beads, the CD4+ specific beads target only your T cells. The whole solution is passed through a column with buffer, but the column is attached to a strong magnet. After a few wash steps you elute the beads (away from magnet ofcourse) and detach the beads from the cells.

https://en.wikipedia.org/wiki/Magnetic-activated_cell_sorting

I agree with Tim that you could separate T cells from other cell types using MACS. I would however recommend bead based methods that don't involve the use of columns. These are much faster (10-20 mins) and less stressful on the cells.

Both BioLegend (Mojosort) and Stemcell Technologies (Easysep) offer T cell isolation kits (negative selection for untouched T cells) and reagents for selective isolation of T cells (excellent purity and yield). The magnets are interchangeable between the two brands.

Hi Suman.... I agree with both Tim and Julie.... However, for downstream applications... negatively sorted cells are better in my experience.... If you have option, you can add biotin labelled antibody against K562 cells in your co-culture, followed by addition of Streptavidin magnetic beads..... Now, when you will place these cells in magnet, you will get negatively selected (untouched) CD4+ cells...

Good luck

Hi all,

A extension of the same question:

If the K562 cells form aggregates with the T cells, how do you separate the cells from each other?