T-Cell Enrichment: A Technique to Isolate T-Cells from Mixed Cell Population by Magnetic Separation

- T-cells, a class of lymphocytes, play an active role in fighting infection. Along with multiple other blood cell types, immunocompetent T-cells are present in mouse spleen and lymph nodes. To selectively isolate T-cells, harvest murine spleen and lymph nodes and macerate them on a cell strainer. Flush the tissue with an appropriate media to obtain a single cell suspension.

Add ammonium-chloride potassium, or ACK buffer, to lyse the red blood cells. Centrifuge to separate the intact blood cells from the lysate. Discard the supernatant and resuspend the cells in a suitable buffer. Treat the suspension with a mixture of biotinylated antimouse antibodies to target blood cells like granulocytes, B-cells, NK-cells, and any remaining erythrocytes. T-cells are not targeted and hence remain untagged.

Next, add anti-biotin magnetic microbeads to the suspension. All biotin-labeled cells bind to the microbeads. When kept under the influence of a strong magnetic field, the magnetically labeled cells adhere to the tube wall. Unlabeled T-cells remain in suspension. Collect the unbound enriched T-cells into a fresh collection tube. In the following protocol, we show T-cells' isolation from mouse