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Immortalized p19 ARF null hepatocytes restore liver injury and generate hepatic progenitors after transplantation
Author(s) -
Mikula Mario,
Fuchs Eva,
Huber Heidemarie,
Beug Hartmut,
SchulteHermann Rolf,
Mikulits Wolfgang
Publication year - 2004
Publication title -
hepatology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.488
H-Index - 361
eISSN - 1527-3350
pISSN - 0270-9139
DOI - 10.1002/hep.20084
Subject(s) - progenitor cell , hepatocyte , biology , transplantation , apoptosis , microbiology and biotechnology , cancer research , liver transplantation , cell culture , transfection , stem cell , immunology , in vitro , medicine , genetics
Primary hepatocytes are blocked in mitotic activity and well‐defined culture conditions only allow the limited expansion of these cells. Various genetic modifications have therefore been employed to establish immortalized hepatic cell lines, but, unfortunately, proper hepatocyte cultures conducting a faithful hepatic gene expression program and lacking malignancy are hardly available. Here we report the immortalization of primary hepatocytes isolated from p19 ARF null mice using the rationale that loss of p19 ARF lowers growth‐suppressive functions of p53 and bypasses cellular senescence without losing genetic stability. The established hepatocytes, called MIM, express liver‐specific markers, show a nontumorigenic phenotype, and competence to undergo Fas‐mediated apoptosis. Intrasplenic transplantation of GFP‐expressing parental MIM cells into Fas‐injured livers of SCID mice revealed liver‐reconstituting activity. In the regenerated liver, MIM cells localized in small‐sized clusters and showed presence in structures comparable to canals of Hering, the site of oval cells. Transplantation of MIM‐Bcl‐X L cells, which are protected against apoptosis, and successive Fas‐induced liver damage, enhanced donor‐derived liver repopulation by showing differentiation into cholangiocytes and cells expressing markers characteristic of both fetal hepatocytes and oval cells. In conclusion, these data indicate that long‐term cultivated p19 ARF null hepatocytes are capable of generating hepatic progenitor cells during liver restoration, and thus represent a highly valuable tool to study the differentiation repertoire of hepatocytes. (H EPATOLOGY 2004;39:628–634.)