Description: The Immortalized Mouse Hepatic Stellate Cells were derived from TLR4-/- mouse via enzymatic digestion and Percoll density gradient centrifugation. The primary cells were immortalized via transfection of the pCMV Simian Virus 40 Large T antigen and hygromycin B selection. The Immortalized Mouse TLR4-/- Hepatic Stellate Cells are useful in studies focusing on liver fibrosis, especially in the role of the pro-inflammatory TLR4 activation in scar tissue formation. TLR4 lentivirus-mediated knock-in experiments have been performed on Plenti4/TO/V5-DEST GatewayTM vectors (Invitrogen) and lipofectamineTM 2000 (Thermo Fisher Scientific). The cells can be used in studies of cellular survival regulation, as TLR4 activation has been linked to apoptosis-resistance.Immortalization Method: Transfection of CMV Simian Virus 40 Large T antigenBioSafety Level: IISpecies: MouseSource Organ: LiverGrowth Properties: AdherentMorphology: PolygonalPopulation Doubling: N/ASeeding Density: N/AMarkers: N/ADonor Age: N/ADonor 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: 33.0°C.
* abm does not recommend to use heat-inactivated FBS for cell culture unless specified otherwise.Quality Control: 1) Genotype analysis; 2)NF-κB Activation Assay; 3) Cytokine ELISA Assays; 4) 3[H]-thymidine Cell Proliferation AssayShipping Condition: Dry IceStorage Condition: liquid nitrogen or -180°C