Abstract
Separation of pure cell populations from the liver is a prerequisite to study the role of hepatic parenchymal and non-parenchymal cells in liver physiology, pathophysiology, and immunology. Traditional methods for hepatic cell separation usually purify only single cell types from liver specimens. Here, we describe an efficient method that can simultaneously purify populations of hepatocytes (HCs), liver sinusoidal endothelial cells (LSECs), and Kupffer cells (KCs) from a single mouse liver specimen. A liberase-based perfusion technique in combination with a low-speed centrifugation and magnetic-activated cell sorting (MACS) led to the isolation and purification of HCs, KCs, and LSECs with high yields and purity.
Key words
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Kmiec Z (2001) Cooperation of liver cells in health and disease. Adv Anat Embryol Cell Biol 161:III–XIII, 1–151
Alpini G, Phillips JO, Vroman B, LaRusso NF (1994) Recent advances in the isolation of liver cells. Hepatology 20(2):494–514
Liu J, Jiang M, Ma Z, Dietze KK, Zelinskyy G, Yang D, Dittmer U, Schlaak JF, Roggendorf M, Lu M (2013) TLR1/2 ligand-stimulated mouse liver endothelial cells secrete IL-12 and trigger CD8+ T cell immunity in vitro. J Immunol 191(12):6178–6190
Seglen PO (1976) Preparation of isolated rat liver cells. Methods Cell Biol 13:29–83
Alabraba EB, Curbishley SM, Lai WK, Wigmore SJ, Adams DH, Afford SC (2007) A new approach to isolation and culture of human Kupffer cells. J Immunol Methods 326(1–2):139–144
Braet F, De Zanger R, Sasaoki T, Baekeland M, Janssens P, Smedsrod B, Wisse E (1994) Assessment of a method of isolation, purification, and cultivation of rat liver sinusoidal endothelial cells. Lab Invest 70(6):944–952
Bhogal RH, Hodson J, Bartlett DC, Weston CJ, Curbishley SM, Haughton E, Williams KT, Reynolds GM, Newsome PN, Adams DH, Afford SC (2011) Isolation of primary human hepatocytes from normal and diseased liver tissue: a one hundred liver experience. PLoS One 6(3):e18222
Chang W, Yang M, Song L, Shen K, Wang H, Gao X, Li M, Niu W, Qin X (2014) Isolation and culture of hepatic stellate cells from mouse liver. Acta Biochim Biophys Sin (Shanghai) 46(4):291–298
Irving MG, Roll FJ, Huang S, Bissell DM (1984) Characterization and culture of sinusoidal endothelium from normal rat liver: lipoprotein uptake and collagen phenotype. Gastroenterology 87(6):1233–1247
Valatas V, Xidakis C, Roumpaki H, Kolios G, Kouroumalis EA (2003) Isolation of rat Kupffer cells: a combined methodology for highly purified primary cultures. Cell Biol Int 27(1):67–73
Daneker GW, Lund SA, Caughman SW, Swerlick RA, Fischer AH, Staley CA, Ades EW (1998) Culture and characterization of sinusoidal endothelial cells isolated from human liver. In Vitro Cell Dev Biol Anim 34(5):370–377
Friedman SL, Roll FJ (1987) Isolation and culture of hepatic lipocytes, Kupffer cells, and sinusoidal endothelial cells by density gradient centrifugation with Stractan. Anal Biochem 161(1):207–218
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2017 Springer Science+Business Media LLC
About this protocol
Cite this protocol
Liu, J., Huang, X., Werner, M., Broering, R., Yang, D., Lu, M. (2017). Advanced Method for Isolation of Mouse Hepatocytes, Liver Sinusoidal Endothelial Cells, and Kupffer Cells. In: Guo, H., Cuconati, A. (eds) Hepatitis B Virus. Methods in Molecular Biology, vol 1540. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6700-1_21
Download citation
DOI: https://doi.org/10.1007/978-1-4939-6700-1_21
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-6698-1
Online ISBN: 978-1-4939-6700-1
eBook Packages: Springer Protocols