An optimized protocol for isolation of hepatic leukocytes retrieved from murine and NASH liver biopsies

一种从鼠类和非酒精性脂肪性肝炎(NASH)肝脏活检组织中分离肝脏白细胞的优化方案

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作者:Ehsan Bahrami ,Tobias Geiger ,Agnes A Steixner-Kumar ,Diana Santacruz ,Coralie Viollet ,Alec Dick ,Yvonne Roth ,Patrycja Schlingeloff ,Julian Schmidberger ,Mark Haenle ,Wolfgang Kratzer ,Kerstin Kitt ,Heike Neubauer ,Eric Simon ,Oliver Krenkel ,Markus Werner

Abstract

Immune dysregulation and inflammation by hepatic-resident leukocytes is considered a key step in disease progression of non-alcoholic fatty liver disease and non-alcoholic steatohepatitis toward cirrhosis and hepatocellular carcinoma. Here, we provide a protocol for isolation and characterization of liver-resident immune cells from fine-needle biopsies obtained from a rodent model and humans. We describe steps for isolating leukocytes, cell sorting, and RNA extraction and sequencing. We then detail procedures for low-input mRNA sequencing analyses. Keywords: Cell Isolation; Clinical Protocol; Flow Cytometry/Mass Cytometry; Health Sciences; Immunology; Molecular Biology; RNA-seq; Sequencing.

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