Characterization of liver function in transdifferentiated hepatocytes

✍ Scribed by Zoë D. Burke; Chia-Ning Shen; Kate L. Ralphs; David Tosh

• Publisher: John Wiley and Sons
• Year: 2005
• Tongue: English
• Weight: 999 KB
• Volume: 206
• Category: Article
• ISSN: 0021-9541
• DOI: 10.1002/jcp.20438

✦ Synopsis

Abstract

We previously demonstrated that dexamethasone (Dex) induces the transdifferentiation (or conversion) of the pancreatic progenitor cell line AR42J‐B13 (B13) to hepatocytes based on the expression of liver proteins. We have extended our original observations to determine: (1) the effects of Dex on pancreatic gene expression; (2) the time course of expression of liver enriched transcription factors during conversion from pancreatic to hepatic phenotype; (3) the functional potential of transdifferentiated hepatocytes; (4) the proliferative capacity of transdifferentiated hepatocytes; and (5) whether ectopic expression of transcription factors can induce the hepatic phenotype in pancreatic B13 cells. The results were as follows. The B13 cell markers amylase, synaptophysin, and neurofilament were lost in transdifferentiated hepatocytes compared to control cells and the liver enriched transcription factors C/EBPβ and C/EBPα were induced first, followed by HNF4α and then RXRα. Using RT‐PCR analysis and immunolocalisation studies, we detected hepatic markers (e.g., apolipoprotein B) in Dex‐treated cells. In transdifferentiated hepatocytes albumin was secreted, insulin stimulated lipid deposition and ciprofibrate enhanced the expression of catalase. Proliferation of transdifferentiated hepatocytes is promoted in the presence of HGF and NEAA as indicated by the co‐expression of the cell cycle markers cyclin D and phosphohistone H3 with liver proteins. Lastly, ectopic expression of C/EBPα or C/EBPβ in AR42J‐B13 cells was sufficient to induce transdifferentiation, based on nuclear localization of HNF4α and induction of UDP‐glucuronosyltransferase expression. These results indicate that the B13 progenitor cell model is suitable for studying liver function and for understanding the molecular and cellular events that occur during transdifferentiation. © 2005 Wiley‐Liss, Inc.