Kupffer Cell Isolation
Kupffer Cell Isolation - Related Videos
03:53
Isolation and Culture of Mouse Kupffer Cells
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2025
In this video, we demonstrate the isolation of Kupffer cells or liver macrophages from the mouse liver. The liver is perfused with a digestion solution followed by density gradient separation. The isolated adherent Kupffer cells can be used for further analysis.
09:49
Kupffer Cell Isolation for Nanoparticle Toxicity Testing
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Cited by 28 •
2015
Liver macrophages, named Kupffer cells, are responsible for the capture of circulating nanoparticles. We describe here a method, of high cell purity and yield, for Kupffer cell isolation. The modified LDH assay is used here to measure the toxicity induced by carbon nanotubes in Kupffer cells.
10:25
"Liver-on-a-Chip" Cultures of Primary Hepatocytes and Kupffer Cells for Hepatitis B Virus Infection
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Cited by 23 •
2019
The goal of this protocol is to provide a step-by-step guide to perform 3-D "liver-on-a-chip" infection experiments with the hepatitis B virus.
08:11
Analysis of Gene Function and Visualization of Cilia-Generated Fluid Flow in Kupffer's Vesicle
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Cited by 19 •
2013
Cilia-generated fluid flow in Kupffer’s Vesicle (KV) controls left-right patterning of the zebrafish embryo. Here, we describe a technique to modulate gene function specifically in KV cells. In addition, we show how to deliver fluorescent beads into KV to visualize fluid flow.
07:26
Isolation of Functional Cardiac Immune Cells
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Cited by 9 •
2011
This method for isolating functional immune cells from the heart provides an alternative to the conventional methods of collagenase digestion, which causes unwanted immune cell activation, resulting in a decreased responsiveness of these cells. Our method of isolation yields functional cardiac immune cells by avoiding problems associated with enzymatic digestion.
FAQs
Perfusion removes blood from the liver before tissue processing, helping reduce blood-derived cellular material in the preparation. This creates a cleaner starting sample for subsequent enzymatic digestion and enrichment of nonparenchymal cells. Because the goal is to study resident hepatic macrophages, reducing blood contamination improves the relevance of later analyses of phagocytosis, cytokine production, and toxicant responses.
Enzymatic digestion releases cells from the liver’s tissue matrix, producing a mixed cellular suspension rather than directly yielding purified Kupffer cells. Differential centrifugation or density-gradient separation then enriches nonparenchymal cells, the group containing the target population. Selective adherence or marker-based purification can provide an additional separation stage when experiments require a more focused Kupffer cell preparation.
Enrichment increases the proportion of Kupffer cells or other nonparenchymal cells in a sample, whereas purification applies further selection to obtain a more specific population. Differential centrifugation and density-gradient separation serve primarily as enrichment approaches. Selective adherence or marker-based purification can then narrow the preparation, allowing investigators to interpret macrophage-specific responses with greater focus.
The preparation depends on how thoroughly the liver is perfused, how effectively enzymes release cells from the tissue matrix, and which separation strategy follows digestion. Differential centrifugation or density gradients may yield an enriched nonparenchymal fraction, while adherence or marker-based selection changes the degree of cellular specificity. These choices should match whether the study requires enrichment or focused macrophage analysis.
A typical workflow begins with liver perfusion, followed by enzymatic digestion to release cells from the tissue matrix. The resulting suspension undergoes differential centrifugation or density-gradient separation to enrich nonparenchymal cells. Researchers may then use selective adherence or marker-based purification before examining the preparation for phagocytosis, cytokine production, toxicant responses, or cellular interactions.
Isolated Kupffer cells are useful when investigators need to examine hepatic immune and inflammatory functions directly. Experiments can assess phagocytosis, cytokine production, responses to toxicants, or interactions with hepatocytes and other immune cells. The approach supports research on liver injury, infection, fibrosis, drug safety, and macrophage biology by connecting cellular behavior with specific liver-related conditions.