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J Immunol Methods. 1990 Aug 28;132(1):137-44.

Isolation and flow cytometric analysis of the free lymphomyeloid cells present in murine liver.

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1
Unité d'Immunophysiologie Cellulaire de l'Institut Pasteur et du C.N.R.S., Paris, France.

Abstract

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 ratio of 2.8), B lymphocytes (20% +/- 2 IgG and 30% IgM positive) and myelomonocytic cells (14% +/- 2 complement receptor type III positive).

PMID:
2202764
DOI:
10.1016/0022-1759(90)90407-m
[Indexed for MEDLINE]

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