Open AccessAn improved experimental method for simultaneously isolating hepatocytes and hepatic stellate cells in mouse liver infected with Echinococcus multilocularis
Author(s) -
Bi Xiaojuan,
Yang Ning,
Ke Ying,
Liu Hui,
Ma Wenmei,
Fang Bingbing,
Sun Li,
Li Liang,
Lü Guodong,
Lin Renyong
Publication year - 2021
Publication title -
journal of clinical laboratory analysis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.536
H-Index - 50
eISSN - 1098-2825
pISSN - 0887-8013
DOI - 10.1002/jcla.24084
Subject(s) - echinococcus multilocularis , hepatic stellate cell , hepatocyte , biology , pathology , cytoplasm , desmin , collagenase , microbiology and biotechnology , echinococcosis , immunohistochemistry , immunology , in vitro , biochemistry , vimentin , medicine , endocrinology , enzyme
Abstract Background Alveolar echinococcosis (AE) is a zoonotic disease caused by the larval stage of Echinococcus multilocularis parasitizing in the human liver, causing local pathological changes in the liver and manifesting as hyperplasia, liver fibrosis, atrophy, degeneration, and necrosis. Here, we report a method that can simultaneously isolate hepatocytes and hepatic stellate cells (HSCs) from mice infected with Echinococcus multilocularis . Methods A mouse model of AE was established. Hepatocytes and HSCs were isolated from mouse liver using a two‐step method combining in situ collagenase perfusion and gradient centrifugation. Expressions of Alb, Desmin, and α‐SMA were detected with immunofluorescence to identify the isolated hepatocytes and HSCs. Results The viability and purity of hepatocytes and HSCs both reached 90% or above. For hepatocytes, clear cell boundaries were observed, and the nuclei were round or oval, with clear nucleoli. There was a homogeneous distribution of the hepatocyte marker Alb in the cytoplasm of hepatocytes. Lipid droplets and Desmin expression were observed in the cytoplasm of freshly isolated HSCs. During the activation of HSCs, the lipid droplets gradually decreased and disappeared with a high expression of α‐SMA. Conclusion Hepatocytes and HSCs are simultaneously isolated. This may provide a research tool to investigate the interaction between hepatocytes and HSCs and to investigate the mechanism of Echinococcus multilocularis infection‐induced liver pathological changes.