Recovery of mature hepatocytic phenotype following bile ductular transdifferentiation of rat hepatocytes in vitro
著者
Sone, Masayuki
Nishikawa, Yuji
Nagahama, Yasuharu
Kumagai, Eriko
Doi, Yuko
Omori, Yasufumi
Yoshioka, Toshiaki
Tokairin, Takuo
Yoshida, Masayuki
Sugiyama, Toshihiro
Enomoto, Katsuhiko
上位タイトル
The American journal of pathology
Vol.181,
No.6
(2012.
12)
,p.2094-
2104
識別番号
ISSN
0002-9440
DOI
10.1016/j.ajpath.2012.08.034
抄録
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.
