The Immortalized Rat Conjunctival Epithelial Cells (CJ4.3) were derived from primary conjunctival epithelial cells from Fischer 344 rats. The primary cells were immortalized with pSV(neo) and transfected with SV40 large T antigen. The immortalized cells display epithelial morphology and express the conjunctiva-specific cytokeratin 4, the goblet cell-specific cytokeratin 7 and were negative for the corneal epithelial cell-specific cytokeratin 12. The Immortalized Rat Conjunctival Epithelial Cells (CJ4.3) is useful as models for studying the conjunctival epithelial barrier and also in developing strategies for delivering macromolecular antigens across this barrier to underlying components of the immune system.
Immortalization Method: Immortalization by transfection with SV40 large T antigen
BioSafety Level: II
Species: Rat
Source Organ: Eye
Growth Properties: Adherent
Morphology: Epithelial-like
Population Doubling: N/A
Seeding Density: Thaw entire contents into an appropriate T25 flask as specified in the Propagation instructions.
Markers: Cytokeratin 4, Cytokeratin 7
Donor Age: N/A
Donor Gender: N/A
Donor Ethnicity: N/A
Propagation: The base medium for this cell line is Prigrow IV medium available at abm (TM004). To make the completed growth medium, add the following components to the base medium: BSA to final concentration of 0.5%, 10 mM HEPES, 1X trace elements B, 10 μg/ml insulin, 5.5 μg/ml transferrin, 50 ng/ml sodium selenate, 20 μg/ml ethanolamine, 0.3 μg/ml glucagon, 0.1 μM dexamethasone, 0.1 μM retinoic acid, 0.3 μM 3, 3’, 5 triiodo-L thyronine, 80 ng/ml epidermal growth factor, 50 μg/ml gentamycin sulfate (Invitrogen-GIBCO) and Penicillin/Streptomycin Solution (G255) to a final concentration of 1%. Carbon dioxide (CO2): 5%, Temperature: +37.0°C.
Quality Control: 1) Immunofluorescence confocal microscopy demonstrating cytokeratin expression
Shipping Condition: Dry Ice
Storage Condition: liquid nitrogen or -180°C