Immortalized Rat Portal Myofibroblasts (RGF)

Activated primary rat portal fibroblasts (PF) were immortalized with the SV40 large T antigen mammalian expression vector to generate the Immortalized Rat Portal Myofibroblasts (RGF). RGF cells are negative for hepatic stellate cell markers, and like the primary cells, they 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 offers the sorted Immortalized Rat Portal Myofibroblasts (RGF-N2) (T0069).

Immortalization Method: Serical passaging and chemical mediated transfection with SV40 large T Antigen mammalian expression vector

BioSafety Level: II

Species: Rat

Source Organ: Liver

Growth Properties: Adherent

Morphology: Spindle shaped

Population Doubling: 13 - 23 hours

Seeding Density: 15,000 - 20,000 cells/cm²

Markers: alpha smooth muscle actin, type I collagen alpha-1, tissue inhibitor of metalloproteinases-1, PF-specific markers elastin, type XV collagen alpha-1, Ntpdase2/Cd39l1, and ecto-5’-nucleotidase/Cd73

Donor Age: Adult

Donor Gender: N/A

Donor Ethnicity: N/A

Propagation: 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 Ice

Storage Condition: liquid nitrogen or -180°C