Journal article

Publication year: 2022

Journal: Biomedicines

Volume number: 10

Issue number: 9

eISSN: 2227-9059

Open access status: Gold

DOI Link: https://doi.org/10.3390/biomedicines10092198

Publisher: MDPI

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 degrees 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.

Harvard Citation style: Poggel, C., Adams, T., Janzen, R., Hofmann, A., Hardt, O., Roeb, E., et al. (2022) Isolation of Hepatocytes from Liver Tissue by a Novel, Semi-Automated Perfusion Technology, Biomedicines, 10(9), Article 2198. https://doi.org/10.3390/biomedicines10092198

APA Citation style: Poggel, C., Adams, T., Janzen, R., Hofmann, A., Hardt, O., Roeb, E., Schroeder, S., Tag, C., Roderfeld, M., & Weiskirchen, R. (2022). Isolation of Hepatocytes from Liver Tissue by a Novel, Semi-Automated Perfusion Technology. Biomedicines. 10(9), Article 2198. https://doi.org/10.3390/biomedicines10092198