Immortalized Slc4a11<sup>-/-</sup> Mouse Corneal Endothelial Cells | T0699
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abm | Immortalized Slc4a11-/- Mouse Corneal Endothelial Cells | T0699
Intercrossed the Slc4a11þ/þ and Slc4a11/C57BL/6 mice with H-2Kb-tsA58 Immortomouse, which carries a temperature-sensitive mutant form of the simian virus-40 large T antigen
Use of FNC coating is required for cell adhesion to the culture vessels. Grow cells in FNC-coated culture vessels with the following conditions.
The base medium for this cell line is Opti-MEM I (Invitrogen). To make the complete growth medium, add the following components to the base medium: heat-inactivated fetal bovine serum to a final concentration of 8%, Antibiotic/Antimycotic to a final concentration of 1%, chondroitin sulfate (Sigma) to a final concentration of 0.08%, CaCl2 (Sigma) to a final concentration of 200 μg/mL, pituitary extract (Biomedical Technologies. Inc.) to a final concentration of 100 μg/mL, gentamycin (Invitrogen) to a final concentration of 0.5%, EGF to a final concentration of 5 ng/ml (Millipore), and IFN-γ (R&D Systems) to a final concentration of 200 ng/mL.
Change media every other day.
Carbon dioxide (CO2): 5%, Temperature: 33.0°C.
Growing cells at 37.0°C will decrease tsTAg levels and stop replication.
1) Western blot; 2) Intracellular pH (pHi) measurement; 3) RT-PCR; 4) Real-time quantitative PCR
liquid nitrogen or -180C
1x10⁶ cells / 1.0 ml