Liver Digestion Protocol
Liver Digestion Protocol - Related Videos
08:47
Protocol for Isolation of Primary Human Hepatocytes and Corresponding Major Populations of Non-parenchymal Liver Cells
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Cited by 96 •
2016
A technique to isolate human hepatocytes and non-parenchymal liver cells from the same donor is described. The different liver cell types build the basis for functional liver models and tissue engineering. This new method aims to isolate liver cells in a high yield and viability.
09:03
Trypsin Digest Protocol to Analyze the Retinal Vasculature of a Mouse Model
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Cited by 59 •
2013
Trypsin digest is one of the most commonly used methods to analyze retinal vasculature. This manuscript describes the method in detail, including key alterations to optimize the technique and remove the non-vascular tissue while preserving the overall architecture of the vessels.
12:35
In vivo Liver Endocytosis Followed by Purification of Liver Cells by Liver Perfusion
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Cited by 14 •
2011
The study of liver sinusoidal endothelial cells (SECs) must be performed with primary cells obtained from the animal as no cell lines exist. This method relies on liver digestion and differential centrifugation for SEC purification for subsequent culturing and experimentation.
08:07
Digestion of the Murine Liver for a Flow Cytometric Analysis of Lymphatic Endothelial Cells
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Cited by 15 •
2019
The goal of this protocol is to identify lymphatic endothelial cell populations within the liver using described markers. We utilize collagenase IV and DNase and a gentle mincing of tissue, combined with flow cytometry, to identify a distinct population of lymphatic endothelial cells.
09:45
Restriction Enzyme Digests
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2023
Restriction enzymes or endonucleases recognize and cut DNA at a specific sequence. These enzymes occur naturally in bacteria as a defense against bacteriophages - viruses that infect bacteria. Bacterial restriction enzymes cut the invading bacteriophage DNA while leaving the bacterial genomic DNA unharmed due to addition of methyl groups. This video explains the basic principles of restriction enzymes including: how restriction enzymes are named and the types of recognition sites and...
FAQs
Proteases weaken cell-cell connections, while collagen-degrading enzymes break down the extracellular matrix that holds liver tissue together. Their combined action releases hepatocytes, immune cells, and other hepatic populations into a suspension suitable for analysis. Balancing these activities is important because insufficient digestion can leave tissue incompletely dissociated, whereas excessive enzymatic exposure may reduce the quality of recovered cells.
Temperature and exposure time control the balance between tissue dissociation and preservation of viable cells. Conditions that are too mild may limit release of resident or infiltrating populations, while prolonged or poorly controlled digestion can compromise cell viability. Managing both variables supports a suspension that is more suitable for downstream flow cytometry, cell culture, and immune profiling.
Controlled perfusion and mincing provide different ways to expose liver tissue to the digestion process. Perfusion applies the treatment through the tissue, whereas mincing first reduces tissue into smaller pieces. The selected approach influences how evenly enzymes contact the sample and can affect the consistency of cell release, making preparation conditions important for reproducible hepatic analyses.
The workflow begins with controlled tissue preparation through perfusion or mincing, followed by enzymatic treatment to disrupt extracellular matrix and cell-cell connections. The resulting material is processed as a viable cell suspension for the selected assay. Maintaining appropriate digestion conditions throughout helps preserve the mixed hepatic population needed for cellular, immunological, or infection-focused measurements.
A liver-derived cell suspension can support flow cytometry, cell culture, pathogen-response studies, and immune-cell profiling. Flow cytometry can examine cellular populations, while culture and response studies provide platforms for investigating how recovered cells behave under experimental conditions. Using the same dissociated material across these applications can connect cellular composition with functional responses in liver research.
The method enables analysis of hepatocytes together with resident and infiltrating immune populations from the same tissue context. This supports investigation of hepatic inflammation, host-pathogen interactions, and tissue-specific immune responses. Because the recovered cells remain organized as an experimentally accessible suspension, researchers can examine how infection-related or inflammatory processes involve different cellular populations within the liver.