Immortalized Slc4a11^+/+ Mouse Corneal Endothelial Cells | T0698

abm | Immortalized Slc4a11^+/+ Mouse Corneal Endothelial Cells | T0698

+/+ Mouse Corneal Endothelial Cells were isolated from the cornea of progeny mice produced by the intercrossing of 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. Immortalized Slc4a11^+/+ Mouse Corneal Endothelial Cells also demonstrates temperature sensitive and IFN-c dependent proliferation. This cell line represents a useful model for the investigation of pathology and therapies for SLC4A11 associated congenital hereditary endothelial dystrophy (CHED) and Fuchs’ endothelial corneal dystrophy.

Immortalization Method:

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

BioSafety Level:

II

Organism:

Mouse

Species:

Mouse

Source Organ:

Eye

Organ Type:

Eye

Growth Properties:

Adherent

Morphology:

Fibroblast-like

Passage Number:

N/A

Population Doubling:

18.72 hours

Seeding Density:

N/A

Markers:

N/A

Donor Age:

N/A

Donor Gender:

N/A

Donor Ethnicity:

N/A

Propagation:

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.

Quality Control:

1) Western blot; 2) Intracellular pH (pHi) measurement; 3) RT-PCR; 4) Real-time quantitative PCR

Shipping Condition:

Dry Ice

Storage Condition:

liquid nitrogen or -180C