Isolation of primary human liver cells from normal and nonalcoholic steatohepatitis livers

Authors:
Xiao Liu , Kevin Lam , Huayi Zhao , Sadatsugu Sakane , Hyun Young Kim , Alvaro Eguileor , Karin Diggle , Shuai Wu , Raquel Carvalho Gontijo Weber , Pejman Soroosh , Mojgan Hosseini , Kristin Mekeel , David A Brenner , Tatiana Kisseleva 
In:
Source: STAR Protoc
Publication Date: (2023)
Issue: 4: 3
Research Area:
Gastroenterology
Toxicology
Cells used in publication:
Hepatocyte, human
Species: human
Tissue Origin: liver
Experiment

The composition of NPCs is best characterized using flow cytometry (Figures 5A and 5B). When the composition of non-parenchymal cells from normal and diseased donors was compared (vs. commercially available NPCs from Lonza Bioscience; https://bioscience.lonza.com/) similar cellular subsets were detected. As expected, the number of myeloid cells was increased in NPCs from the diseased donor, while the number of CD3+ T cells was decreased. Although LSECs were tested for expression of CD31, the percent of LSECs was low in both populations (Figure 5C). It is also recommended to test expression of CD146 in LSEC vs. all endothelial cells as expression of CD146 is associated with LSECs. CD146 and others such as Stabilin2 are good markers for LSECs. CD31 also detects endothelial cells from larger blood vessels, and these are not LSECs. Recent studies have suggested that Scavenger receptor type B1(SR-B1) is specifically expressed by LSECs

Abstract

Here, we present a protocol for isolating human hepatocytes and neural progenitor cells from normal and nonalcoholic steatohepatitis livers. We describe steps for perfusion for scaled-up liver cell isolation and optimization of chemical digestion to achieve maximal yield and cell viability. We then detail a liver cell cryopreservation and potential applications, such as the use of human liver cells as a tool to link experimental and translational research.