I processed 4 liver from CD1 mice for lymphocyte isolation. I followed a protocol without perfusion of liver. I got very low lymphocyte count: range from 1-5 million cells only from each liver. Is it normal? I usually get 90-130 million splenocytes from spleen. Since liver is larger than spleen, I thought, I should be even more cells.

My steps were:

1) Harvest the liver and get rid of gall bladder.

2) Cut the liver into small pieces

3) Smashed the liver pieces and pass through 70um strainer

4) 35% Percoll density gradient separation.

5) RBC lysis.

6) Cell count.

Any suggestion to get more lymphocytes from the liver?