Single Cell Suspension From Spleen

Harvest mouse spleen and prepare a single cell suspension.

Single cell suspension from spleen.

Peripheral blood lymph and lymphoid organs. Aim of this experiment is to isolate single cell suspension from the spleen and thymus of the mouse. Resuspend the pellet in 5 ml spleen of lysis buffer. This video outlines how to harvest cells from a primary tissue sample and prepare a single cell suspension prior to performing cell isolation.

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 11 set up another cell culture dish with 5 10 ml d pbs. This video demonstrates how to harvest cells from a spleen sample and prepare a single cell suspension prior to performing cell isolation. Pellet the cells by centrifugation 300 400xg at 2 8 c and aspirate the supernatant.

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. 6 12 transfer several tissue pieces to strainer and keep moistened with d pbs. Spleen and thymus are the lymphoid organs of an animal starting from lower vertebrate like fish to higher vertebrate like human. 6 13 open a sterile 3 or 5 ml syringe.

6 10 filter the cell suspension through a sterile cell strainer to remove dispersed cells from tissue fragments. This protocol describes how to harvest cells from a spleen sample and how to prepare a single cell suspension prior to performing cell isolation. Therefore culture of cells derived from these lymphoid organs will help to understand many aspects in the field of immunology. Red blood cell lysis after having prepared the single cell suspension.

Preparing a true single cell suspension of the primary tissue sample will optimize cell separation from tissues by avoiding additional cell loss and enabling maximum labeling of the target cells. I used flow cytometry to detect single cell suspension from mouse spleen and there were too many dead cells. Lysis of mouse erythrocytes red cell lysis. Discard the supernatant and resuspend the cells in pbs w 0 1 bsa to 1 x 10 7 cells ml.

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. The gentlemacs dissociator provides optimized programs to attain single cell suspensions from various tissues for example mouse spleen.