Details for amaxa FAQ article #71

«How do you recommend that we isolate splenic lymphocytes from mouse spleens? Is erythrocyte lysis required?»

For the preparation of mouse spleen cells we recommend cutting the spleen once and passing the tissue through a 100µM cell strainer or steel mesh using a plunger. The cells are flushed into a petri dish containing PBS. In order to remove fat, cell debris and aggregates, the cells can further be passed through a 70µM cell strainer or a cotton wool column. With this procedure, each spleen should yield approximately 50 million-200 million cells. We strongly recommend omitting the erythrocyte lysis step since it leads to a decrease in lymphocyte viability after Nucleofection. Due to relatively low numbers of erythrocytes in the spleen cell preparation (1:2 erythrocytes: leukocytes compared to 1:1000 in human peripheral blood), an erythrocyte lysis is not required.