Isolation of murine hepatic lymphocytes using mechanical dissection for phenotypic and functional analysis of NK1.1+ cells

World Journal of Gastroenterology : WJG
Zhong-Jun DongBin Gao

Abstract

To choose an appropriate methods for the isolation of hepatic lymphocytes between the mechanical dissection and the enzymatic digestion and investigate the effects of two methods on phenotype and function of hepatic lymphocytes. Hepatic lymphocytes were isolated from untreated, poly (I:C)-stimulated or ConA-stimulated mice using the two methods, respectively. The cell yield per liver was evaluated by direct counting under microscope. Effects of digestive enzymes on the surface markers involved in hepatic lymphocytes were represented by relative change rate ((percentage of post-digestion -percentage of pre-digestion)/percentage of pre-digestion). Phenotypic analyses of the subpopulations of hepatic lymphocytes and intracellular cytokines were detected by flow cytometry. The cytotoxicity of NK cells from wild C57BL/6 or poly (I:C)-stimulated C57BL/6 mice was analyzed with a 4-h (51)Cr release assay. NK1.1(+) cell markers, NK1.1 and DX5, were significantly down-expressed after enzymatic digestion and their relative change rates were about 28% and 32%, respectively. Compared with the enzymatic digestion, the cell yield isolated from unstimulated, poly (I:C)-treated or ConA-treated mice by mechanical dissection was not significantly...Continue Reading

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Citations

Aug 7, 2010·European Journal of Immunology·Sandrine TchaptchetMarina A Freudenberg
May 12, 2012·Hepatology : Official Journal of the American Association for the Study of Liver Diseases·Masanobu TsudaM Eric Gershwin
Apr 9, 2013·Hepatology : Official Journal of the American Association for the Study of Liver Diseases·Massimiliano SarraGiovanni Monteleone
Jan 5, 2006·Canadian Journal of Physiology and Pharmacology·Zhongjun DongZhigang Tian
May 5, 2020·American Journal of Physiology. Gastrointestinal and Liver Physiology·Takuro OkamuraMichiaki Fukui
Apr 2, 2021·Frontiers in Immunology·Masahide HamaguchiMichiaki Fukui

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