Immortalized Mouse Hepatocytes-Conditionally (ImHep)

Description: ImHep is a conditionally immortalized mouse hepatocyte isolated from the mouse harboring thermolabile mutation (tsA58) under the control of an interferon (IFN)-γ-inducible H-2K^b promoter and a temperature-sensitive simian virus 40 large T antigen. The functional expression of the SV40 large T antigen is induced by culturing the cells in vitro in medium containing IFN γ at a temperature permissive (33°C). At a non-permissive temperature (37°C-39°C) or when cultured without IFN γ over 3 passages, the cells cease to proliferate. The ImHep cell line is unique in that 1) it is easily transfectable; 2) it is able to persist in vivo post-hepatocllular transplantation at the non-permissive temperature; both of these characteristics make this cell line a valuable tool for in vitro genetic manipulation, and use in mouse models for ex vivo gene therapy.

Immortalization Method: Isolated from transgenic mouse carrying a temperature-sensitive simian virus 40 tumor antigen (tsA58)

BioSafety Level: II

Species: Mouse

Source Organ: Liver

Growth Properties: Adherent

Morphology: Other

Population Doubling: N/A

Seeding Density: Recommended split ratio of 1:3

Markers: albumin, AFP, transferrin, LPK, CK8 and CK18

Donor Age: N/A

Donor Gender: N/A

Donor Ethnicity: N/A

Propagation: The base medium for this cell line is Prigrow II medium available at abm (TM002). To make the completed growth medium, add the following components to the base medium: fetal bovine serum (TM999)* to a final concentration of 5%, 5 U/ml murine IFN-γ, 1 µg/mL insulin, 10^–5 M hydrocortisone and Penicillin/Streptomycin Solution (G255) to a final concentration of 1%.
Carbon dioxide (CO₂): 5%, Temperature: 33.0°C.
* abm does not recommend to use heat-inactivated FBS for cell culture unless specified otherwise.

Quality Control: 1) Western Blot and Immunofluorescence were used to detect hepatocyte markers albumin, AFP, transferrin, LPK, CK8 and CK18; 2) Immunofluorescence were used to verify the presence of SV40 large T antigen

Shipping Condition: Dry Ice

Storage Condition: liquid nitrogen or -180°C