How is a mouse spleen cell preparation carried out?
For the preparation of mouse spleen cells by mechanical disruption, it is recommended to cut the spleen once and to pass the tissue through a steel mesh by using a plunger. Subsequently, the cells are flushed into a petri dish containing PBS. Collagenase treatment is strongly recommended for the isolation of dendritic cells and liver sinusoidal endothelial cells to increase recovery of the cells. In order to remove fat, cell debris and cell aggregates, the cells can be passed through a cotton wool column. With this procedure, each spleen should yield approximately 5×10e7–1×10e8 cells. Due to relatively low numbers of erythrocytes in the spleen cell preparation (2:1 ratio for erythrocytes:leukocytes, compared to a 1000:1 ratio in human peripheral blood), an erythrocyte lysis is not required.
