title: Recovery of mature hepatocytic phenotype following bile ductular transdifferentiation of rat hepatocytes in vitro.
abstract: We previously demonstrated that mature rat hepatocytes transdifferentiate to bile ductular cells when cultured in a three-dimensional collagen-rich matrix. Here, we show that the phenotype of transdifferentiated hepatocytes can be reversed by modulating culture conditions. Spheroidal aggregates of hepatocytes were cultured within a collagen gel matrix in the presence of serum and tumor necrosis factor-α. Spheroids transformed into ductular structures composed of small cuboidal cells, lost the expression of hepatocytic markers, whereas aberrantly expressed bile ductular markers. The transdifferentiated cells were then retrieved from the gels, plated on surfaces coated with a basement membrane-like material, and cultured in serum-free media. Cells spontaneously formed spheroidal aggregates and recovered hepatocytic phenotype. Dexamethasone (Dex), which suppressed the phosphorylation of ERK and Jun N-terminal kinase, facilitated the recovery, and the combination with interleukin-6 or oncostatin M resulted in the recovery of hepatocyte nuclear factor 4 α protein expression and the typical hepatocytic morphology, and a decrease in the expression of bile ductular markers. A cDNA microarray analysis revealed that the hepatocyte-specific mRNA expression profile was recovered in these cells. Our results demonstrate that hepatocytes are able to recover their phenotypes following bile ductular transdifferentiation, suggesting that hepatocytic and bile ductular phenotypes may be mutually reversible.
authors.affiliation: Department of Molecular Pathology and Tumor Pathology, Akita University Graduate School of Medicine, Akita, Japan.
authors.forename: Masayuki
authors.initials: M
authors.lastname: Sone
authors.forename: Yuji
authors.initials: Y
authors.lastname: Nishikawa
authors.forename: Yasuharu
authors.initials: Y
authors.lastname: Nagahama
authors.forename: Eriko
authors.initials: E
authors.lastname: Kumagai
authors.forename: Yuko
authors.initials: Y
authors.lastname: Doi
authors.forename: Yasufumi
authors.initials: Y
authors.lastname: Omori
authors.forename: Toshiaki
authors.initials: T
authors.lastname: Yoshioka
authors.forename: Takuo
authors.initials: T
authors.lastname: Tokairin
authors.forename: Masayuki
authors.initials: M
authors.lastname: Yoshida
authors.forename: Toshihiro
authors.initials: T
authors.lastname: Sugiyama
authors.forename: Katsuhiko
authors.initials: K
authors.lastname: Enomoto
mesh_terms: D000375:Aging; D000818:Animals; D001652:Bile Ducts / Q000166:cytology*; D002469:Cell Separation; D048430:Cell Shape / Q000187:drug effects; D054338:Cell Transdifferentiation / Q000187:drug effects / Q000235:genetics; D003094:Collagen / Q000494:pharmacology; D003907:Dexamethasone / Q000494:pharmacology; D004338:Drug Combinations; D005782:Gels / Q000494:pharmacology; D020869:Gene Expression Profiling; D022781:Hepatocytes / Q000166:cytology* / Q000187:drug effects / Q000201:enzymology / Q000648:ultrastructure; D015850:Interleukin-6 / Q000494:pharmacology; D007797:Laminin / Q000494:pharmacology; D048670:MAP Kinase Kinase 4 / Q000037:antagonists & inhibitors / Q000378:metabolism; D020935:MAP Kinase Signaling System / Q000187:drug effects; D008297:Male; D020929:Mitogen-Activated Protein Kinase Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism; D053683:Oncostatin M / Q000494:pharmacology; D010641:Phenotype; D010766:Phosphorylation / Q000187:drug effects; D011509:Proteoglycans / Q000494:pharmacology; D051381:Rats; D011916:Rats, Inbred F344; D055647:Rats, Transgenic; D014157:Transcription Factors / Q000378:metabolism; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
chemical_list: D004338:Drug Combinations; D005782:Gels; D015850:Interleukin-6; D007797:Laminin; D011509:Proteoglycans; D014157:Transcription Factors; D014409:Tumor Necrosis Factor-alpha; D053683:Oncostatin M; C059485:matrigel; D003907:Dexamethasone; D003094:Collagen; D048670:MAP Kinase Kinase 4; D020929:Mitogen-Activated Protein Kinase Kinases