ISOLATION AND FLOW CYTOMETRIC ANALYSIS OF THE FREE LYMPHOMYELOID CELLS PRESENT IN MURINE LIVER

Recruitment of circulating lymphomyeloid cells in the liver during infection often plays a critical role, mediating control or exacerbation of the pathogen growth. This paper describes a simple and rapid technique to recover these lymphomyeloid cells from a normal or an infected liver. After portal perfusion with saline buffer, the liver is gently dissociated on steel screens and the resulting cell population spun in 35% Percoll in 100 IU/ml Calciparine to remove all nuclei and cell debris: the recovery of a pure liver lymphomyeloid cell population is usually achieved in 40-60 min. Phenotypic and functional analysis could then be easily carried out on this cell population. This methodology was applied to normal mouse liver: flow cytometric analysis of the purified free lymphomyeloid cells showed the presence of T lymphocytes (46%±3 with a CD4 CD8 ration of 208), B lymphocytes (20%±2 IgG and 30% IgM positive) and myelomonocytic cells (14%±2 complement receptor type III positive). © 1990.