The Immortalized Mouse Hepatic Stellate Cells were derived from MyD88-/- 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. Myeloid differentiation factor 88 (MyD88) is an adaptor protein downstream of the TLR4 pathway. LPS activation of the Immortalized Mouse MyD88-/- Hepatic Stellate Cells expressing WT TLR4 has shown almost no change in NF-κB activation. The Immortalized Mouse MyD88-/- Hepatic Stellate Cells are useful in studies focusing on liver fibrosis, especially in the role of the inflammatory TLR4 pathway activation in scar tissue formation. In addition, the cells can be used in studies of cellular survival regulation, since TLR4 activation has been linked to apoptosis-resistance.
Immortalization Method: pCMV Simian Virus 40 Large T antigen
BioSafety Level: II
Species: Mouse
Source Organ: Liver
Growth Properties: Adherent
Morphology: Polygonal
Population Doubling: 25-35 hours
Seeding Density: 10,000 cells/cm²
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