Establishment and Characterization of XXXX Portal Myofibroblast Cell Lines.

@entity1603 @entity33305 @entity6930 @entity4116 @entity71317 @entity35 @entity904 @entity10944 @entity201 @entity1575
UNASSIGNED: The major sources of scar-forming myofibroblasts during @entity1575 are activated hepatic stellate cells (HSC) and portal fibroblasts (PF). In contrast to well-characterized HSC, PF remain understudied and poorly defined. This is largely due to the facts that isolation of rodent PF for functional studies is technically challenging and that PF cell lines had not been established. To address this, we have generated two polyclonal portal myofibroblast cell lines, RGF and RGF-N2. RGF and RGF-N2 were established from primary PF isolated from adult @entity35 livers that underwent culture activation and subsequent SV40-mediated immortalization. Specifically, @entity71317 / @entity71317 -sorted primary PF were used to generate the RGF-N2 cell line. Both cell lines were functionally characterized by RT-PCR, immunofluorescence, immunoblot and @entity1603 -based proliferation assay. First, immortalized RGF and RGF-N2 cells are positive for phenotypic myofibroblast markers alpha smooth muscle actin, @entity6930 , PF-specific markers @entity4116 , type XV collagen alpha-1 and @entity71317 / @entity71317 , and mesenchymal cell marker @entity33305 / @entity33305 , while negative for HSC-specific markers @entity10944 and lecithin @entity904 acyltransferase. Second, both RGF and RGF-N2 cell lines are readily transfectable using standard methods. Finally, RGF and RGF-N2 cells attenuate the growth of Mz-ChA-1 @entity201 cells in co-culture, as previously demonstrated for primary PF. Immortalized @entity35 portal myofibroblast RGF and RGF-N2 cell lines express typical markers of activated PF-derived myofibroblasts, are suitable for DNA transfection, and can effectively inhibit cholangiocyte proliferation. Both RGF and RGF-N2 cell lines represent novel in vitro cellular models for the functional studies of portal (myo)fibroblasts and their contribution to the progression of @entity1575 .
