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

Autor: 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
Jazyk: angličtina
Rok vydání: 2023
Předmět:
Zdroj: STAR Protocols, Vol 4, Iss 4, Pp 102597- (2023)
Druh dokumentu: article
ISSN: 2666-1667
DOI: 10.1016/j.xpro.2023.102597
Popis: Summary: 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. : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.
Databáze: Directory of Open Access Journals