Open AccessFive‐in‐One: Simultaneous isolation of multiple major liver cell types from livers of normal and NASH mice
Author(s) -
Zhou Ye,
Adewale Funmilola,
Kim Sun,
Su Qi,
Glass David,
Sleeman Mark W.,
Murphy Andrew J.,
Cheng Xiping
Publication year - 2021
Publication title -
journal of cellular and molecular medicine
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.44
H-Index - 130
eISSN - 1582-4934
pISSN - 1582-1838
DOI - 10.1111/jcmm.16933
Subject(s) - cell , hepatic stellate cell , liver cell , cell type , cell sorting , chronic liver disease , biology , differential centrifugation , immune system , liver disease , antibody , computational biology , microbiology and biotechnology , immunology , medicine , biochemistry , cirrhosis , endocrinology
NASH is a chronic liver disease that affects 3%–6% of individuals and requires urgent therapeutic developments. Isolating the key cell types in the liver is a necessary step towards understanding their function and roles in disease pathogenesis. However, traditional isolation methods through gradient centrifugation can only collect one or a few cell types simultaneously and pose technical difficulties when applied to NASH livers. Taking advantage of identified cell surface markers from liver single‐cell RNAseq, here we established the combination of gradient centrifugation and antibody‐based cell sorting techniques to isolate five key liver cell types (hepatocytes, endothelial cells, stellate cells, macrophages and other immune cells) from a single mouse liver. This method yielded high purity of each cell type from healthy and NASH livers. Our five‐in‐one protocol simultaneously isolates key liver cell types with high purity under normal and NASH conditions, enabling for systematic and accurate exploratory experiments such as RNA sequencing.