ArticleBiomedicines2022
Isolation of Hepatocytes from Liver Tissue by a Novel, Semi-Automated Perfusion Technology.
Article in Biomedicines, 2022. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 8 papers.
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Who cites it
8 citing papers in PubMed.
- Optimization of Sample Processing for Droplet Digital PCR Quantification ofPathogens (Basel, Switzerland) · 2026Article
- SUCNR1 coordinates metabolic flux, mitochondrial function, and nutrient-dependent adaptation in hepatocytes.Science advances · 2026Article
- Article
- Targeting Oxalate Production by Combining Enzyme Inhibition and Proteolysis Activation: A Novel Therapeutic Approach for Primary Hyperoxaluria Type 1.Journal of medicinal chemistry · 2026Article
- The In Vitro Transgenic Rodent Assay in Primary MutaMouse Hepatocytes Compared to the Mammalian Cell Gene Mutation Assay Using the HPRT Gene.Environmental and molecular mutagenesis · 2026Article
- Optimized Protocol for Primary Rat Hepatocyte Isolation and a Model for Investigating Experimental Steatosis.Methods and protocols · 2025Article
- SPTLC3 regulates plasma membrane sphingolipid composition to facilitate hepatic gluconeogenesis.Cell reports · 2024Article
- Article
Corrections and comments
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Authors and funding
10 authors.
Funding
Abstract
Primary hepatocytes are a major tool in biomedical research. However, obtaining high yields of variable hepatocytes is technically challenging. Most protocols rely on the two-step collagenase perfusion protocol introduced by Per Ottar Seglen in 1976. In this procedure, the liver is perfused in situ with a recirculating, constant volume of calcium-free buffer, which is maintained at 37 °C and continuously oxygenated. In a second step, the liver is removed from the carcass and perfused with a collagenase solution in order to dissociate the extracellular matrix of the liver and liberate individual cells. Finally, the dissected hepatocytes are further purified and concentrated by density-based centrifugation. However, failure in proper cannulation, incomplete enzymatic digestion or over-digestion can result in low cell yield and viability. Here we present a novel semi-automated perfusion device, which allows gentle, rapid and efficient generation of a single-cell suspension from rodent livers. In combination with prefabricated buffers, the system allows reliable and highly reproducible isolation of primary hepatocytes.
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Registered trials
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