Protocol for optimized mononuclear cell isolation from liver and tumor tissue using mechanical or enzymatic digestion

Area of Science:

• * Hepatobiliary surgery
• * Immunology
• * Cancer research

Background:

• * Understanding the tumor microenvironment (TME) in hepatocellular carcinoma (HCC) is crucial for developing effective cancer therapies.
• * Mononuclear cells within liver tissue, encompassing both tumor and non-tumor regions, are key components of the TME.
• * Isolation of these cells is essential for detailed immunological studies.

Purpose of the Study:

• * To present a detailed protocol for the dissociation of human liver tissue from HCC patients.
• * To outline methods for generating a single-cell suspension suitable for downstream analysis.
• * To describe the isolation of mononuclear cells and subsequent immune cell characterization via flow cytometry.

Main Methods:

• * Tissue dissociation using mechanical or enzymatic digestion methods.
• * Separation of mononuclear cells via density gradient centrifugation.
• * Flow cytometry staining for immune cell identification and phenotyping.

Main Results:

• * Successful generation of single-cell suspensions from HCC liver tissue.
• * Efficient isolation of mononuclear cells from both tumor and non-tumor hepatic regions.
• * Demonstrated feasibility of flow cytometry staining for analyzing immune cell populations.

Conclusions:

• * The presented protocol provides a reliable method for isolating mononuclear cells from HCC liver tissue.
• * This technique facilitates the comprehensive study of immune cell infiltrates within the HCC tumor microenvironment.
• * The isolated cells are suitable for advanced immunological analyses, aiding in the development of targeted therapies.