Immortalized Mouse TLR4^-/- Hepatic Stellate Cells

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 Gateway^TM vectors (Invitrogen) and lipofectamine^TM 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 antigen

BioSafety Level: II

Species: Mouse

Source Organ: Liver

Growth Properties: Adherent

Morphology: Polygonal

Population Doubling: N/A

Seeding Density: N/A

Markers: N/A

Donor Age: N/A

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: +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 Assay

Shipping Condition: Dry Ice

Storage Condition: liquid nitrogen or -180°C