Single Cell Suspension From Spleen

Discard the supernatant and resuspend the cells in pbs w 0 1 bsa to 1 x 10 7 cells ml.

Single cell suspension from spleen.

The gentlemacs dissociator provides optimized programs to attain single cell suspensions from various tissues for example mouse spleen. Therefore culture of cells derived from these lymphoid organs will help to understand many aspects in the field of immunology. Preparing a single cell suspension of the primary tissue sample will optimize cell separation by avoiding additional cell loss and enabling maximum labeling of the target cells. 6 10 filter the cell suspension through a sterile cell strainer to remove dispersed cells from tissue fragments.

This video outlines how to harvest cells from a primary tissue sample and prepare a single cell suspension prior to performing cell isolation. I used flow cytometry to detect single cell suspension from mouse spleen and there were too many dead cells. 6 13 open a sterile 3 or 5 ml syringe. This chapter discusses that lymphoid cells may be obtained from various sources.

Spleen and thymus are the lymphoid organs of an animal starting from lower vertebrate like fish to higher vertebrate like human. Continue at step 6 15 below. 6 12 transfer several tissue pieces to strainer and keep moistened with d pbs. Pellet the cells by centrifugation 300 400xg at 2 8 c and aspirate the supernatant.

6 11 set up another cell culture dish with 5 10 ml d pbs. Harvest mouse spleen and prepare a single cell suspension. Lysis of mouse erythrocytes red cell lysis. This video demonstrates how to harvest cells from a spleen sample and prepare a single cell suspension prior to performing cell isolation.

Peripheral blood lymph and lymphoid organs. Aim of this experiment is to isolate single cell suspension from the spleen and thymus of the mouse. Single cell suspensions are a prerequisite for many experiments for example to achieve the highest possible purity and recovery during cell separations with macs technology. Resuspend the pellet in 5 ml spleen of lysis buffer.

Red blood cell lysis after having prepared the single cell suspension. In the case of peripheral blood and lymph the cells are already in single cell suspension and the techniques for the separation of the various populations and subpopulations of cells are also discussed.