QSG-7701 human hepatocytes form polarized acini in three-dimensional culture.
Hepatocytes are polarized and fulfill a variety of liver-specific functions in vivo; but the polarized tissue structure and many of these functions are lost when the cells are cultured on plastic. To recapitulate the polarized structure and tissue-specific function of liver cells in culture, we established a three-dimensional (3D) culture assay with the human hepatocyte line QSG-7701. In 3D Matrigel culture, QSG-7701 cells formed polarized spheroids with a center lumen, which is reminiscent of bile canaliculi in the liver. Immunofluoresence analysis showed that F-actin bundles and radixin were mainly located at the apical membrane and that alpha6 and beta1 integrins were localized basally in 3D culture. Lumen formation was associated with the selective apoptosis of centrally located cells and was accompanied by proliferative suppression during acinar development. Compared to QSG-7701 cells in 2D or agarose gel cultures, the cells in 3D Matrigel culture maintained a given direction of biliary excretion and acquired higher levels of cytochrome P450 and albumin expression. Our study shows that the immortal human hepatocytes, QSG-7701, in 3D Matrigel culture reacquire cardinal features of glandular epithelium in vivo, providing an ex vivo model to study liver-specific function and tumorigenesis.
Zhang
Fang
F
State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China.
Xu
Ren
R
Zhao
Mu-jun
MJ
D000199:Actins / Q000378:metabolism; D015820:Cadherins / Q000378:metabolism; D018929:Cell Culture Techniques / Q000379:methods*; D002460:Cell Line; D016764:Cell Polarity; D049109:Cell Proliferation; D003094:Collagen; D003598:Cytoskeletal Proteins / Q000378:metabolism; D004338:Drug Combinations; D020869:Gene Expression Profiling; D022781:Hepatocytes / Q000166:cytology* / Q000378:metabolism; D006801:Humans; D039503:Integrin alpha6 / Q000378:metabolism; D019012:Integrin beta1 / Q000378:metabolism; D053538:Keratin-18 / Q000235:genetics; D053552:Keratin-7 / Q000235:genetics; D053533:Keratin-8 / Q000235:genetics; D007797:Laminin; D008565:Membrane Proteins / Q000378:metabolism; D008856:Microscopy, Fluorescence; D011509:Proteoglycans; D011518:Proto-Oncogene Proteins / Q000235:genetics; D000074562:Receptor, Notch4; D051880:Receptors, Notch / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D051176:beta Catenin / Q000378:metabolism
D000199:Actins; D015820:Cadherins; D003598:Cytoskeletal Proteins; D004338:Drug Combinations; D039503:Integrin alpha6; D019012:Integrin beta1; D053538:Keratin-18; D053552:Keratin-7; D053533:Keratin-8; D007797:Laminin; D008565:Membrane Proteins; C106075:NOTCH4 protein, human; D011509:Proteoglycans; D011518:Proto-Oncogene Proteins; D000074562:Receptor, Notch4; D051880:Receptors, Notch; D051176:beta Catenin; C059485:matrigel; C071776:radixin; D003094:Collagen