Activated primary rat portal fibroblasts (PF) sorted via the Ntpdase2/Cd39l1 marker were immortalized with the SV40 large T antigen to generate Immortalized Rat Portal Myofibroblasts (RGF-N2). RGF-N2 cells, like primary PFs, demonstrate inhibition of cholangiocyte proliferation in co-culture conditions. The easily transfectable nature of these cells make them an ideal research tool for studying both the function of myofibroblasts and their contribution to liver fibrosis progression.
abm also offers the non sorted Immortalized Rat Portal Myofibroblasts (RGF) (T0068)Immortalization Method: Serical passaging and chemical mediated transfection with SV40 large T Antigen mammalian expression vectorBioSafety Level: IISpecies: RatSource Organ: LiverGrowth Properties: AdherentMorphology: Spindle shapedPopulation Doubling: 17 - 27 hoursSeeding Density: 15,000 - 20,000 cells/cm²Markers: alpha smooth muscle actin, type I collagen alpha-1, tissue inhibitor of metalloproteinases-1, PF-specific markers elastin, type XV collagen alpha-1 and Ntpdase2/Cd39l1, ecto-5’-nucleotidase/Cd73Donor Age: AdultDonor Gender: N/ADonor Ethnicity: N/APropagation: The base medium for this cell line is Prigrow III medium available at abm (TM003). To make the completed growth medium, add the following components to the base medium: fetal bovine serum (TM999)* to final concentration of 10% and Penicillin/Streptomycin Solution (G255) to a final concentration of 1%. Carbon dioxide (CO2): 5%, Temperature: +37.0°C.* abm does not recommend to use heat-inactivated FBS for cell culture unless specified otherwise.Quality Control: 1.) Gene expression analysis to confirm the expression of common liver myofibroblast-associated genes.
2.) Immunoblot and immunofluoresence microscopy to detect myofibroblast-specific cell markers.
3.) Transfection assay to confirm DNA transfectability of cells.
4.) Co-culture assay to demonstrate the inhibition of Mz-ChA-1 cholangiocarcinoma proliferation.Shipping Condition: Dry IceStorage Condition: liquid nitrogen or -180°C