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 ratio of 2.8), B lymphocytes (20% +/- 2 IgG and 30% IgM positive) and myelomonocytic cells (14% +/- 2 complement receptor type III positive).
Unité d'Immunophysiologie Cellulaire de l'Institut Pasteur et du C.N.R.S., Paris, France.
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Goossens
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Jouin
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Marchal
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Milon
D000818:Animals; D000954:Antigens, Surface / Q000032:analysis; D001402:B-Lymphocytes / Q000276:immunology*; D002499:Centrifugation, Density Gradient; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D008099:Liver / Q000166:cytology* / Q000276:immunology; D008297:Male; D051379:Mice; D008809:Mice, Inbred C3H; D008810:Mice, Inbred C57BL; D009000:Monocytes / Q000276:immunology*; D010477:Perfusion; D013601:T-Lymphocytes / Q000276:immunology*
D000954:Antigens, Surface