Isolation and Characterization of Mouse Primary Liver Sinusoidal Endothelial Cells

Area of Science:

• Hepatology
• Cell Biology
• Immunology

Background:

• Liver sinusoidal endothelial cells (LSECs) are crucial for liver function and disease pathogenesis.
• Limited protocols exist for LSEC isolation and characterization, hindering research.
• LSECs are implicated in metabolic disorders, inflammation, fibrosis, and cancer.

Purpose of the Study:

• To provide a detailed protocol for isolating and characterizing LSECs from healthy and nonalcoholic fatty liver disease (NAFLD) mouse models.
• To enable downstream functional and multi-omics analyses of LSECs.

Main Methods:

• Collagenase perfusion of mouse liver followed by magnetic bead-based positive selection of nonparenchymal cells.
• Characterization using flow cytometry for specific markers and scanning electron microscopy for phenotypic features.

Main Results:

• A robust protocol for LSEC isolation from healthy and NAFLD mice.
• Successful characterization of isolated LSECs using flow cytometry and scanning electron microscopy.
• Demonstrated utility of isolated LSECs for functional assays and multi-omics data generation (RNA-seq, proteomics, ATAC-seq).

Conclusions:

• This protocol facilitates in-depth study of LSEC communication and function in liver health and disease.
• Enables comprehensive understanding of LSEC roles in acute and chronic liver injury mechanisms.
• Supports research into LSEC-mediated pathogenesis of liver diseases like NAFLD.