|
Print Version |
| BioSafety Level |
II |
| Organism |
Fischer |
| Source Organ |
Eye |
| Growth Properties |
Adherent |
| Morphology |
Epithelial |
| Recommended Seeding Density |
Thaw entire contents into an appropriate T25 flask as specified in the Propagation instructions. |
| Markers |
Cytokeratin 4, Cytokeratin 7 |
| Applications |
For Research Use Only |
| Immortalization Method |
Immortalization by transfection with SV40 large T antigen |
| Description |
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. |
| Procedure Overview |
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| Image |
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| Propagation |
Use of PriCoat? T25 Flasks (G299) or Applied Cell Extracellular Matrix (G422) is required for cell adhesion to the culture vessels. Grow cells in ECM-coated culture vessels with the following conditions. The base medium for this cell line is PriGrow IV medium available from 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 1% Penicillin/Streptomycin (G255). Atmosphere: air: 95%, CO?: 5%; Temperature: 37.0°C. |
| Subculturing |
1. Remove and discard culture medium.
2. Add 2.0mL of Trypsin-EDTA (TM050) solution to flask and observe cells under an inverted microscope until cell layer is dispersed. Note: Cells that are difficult to detach may be placed at 37°C to facilitate dispersal.
3. Centrifuge cells at 1500rpm for 3 minutes to pellet.
4. Aspirate out trypsin, leaving pellet undisturbed.
5. Resuspend pellet in fresh culture medium and plate in new culture vessel.
6. Incubate cultures at 37°C. Be sure to use T25 ECM-coated flasks(G299) for growth. |
| Preservation |
1. Freeze Medium: Complete growth medium with 20% FBS and 10% DMSO.
2. Storage Temperature: Liquid nitrogen vapour phase. |
| Quality Control |
1) Immunofluorescence confocal microscopy demonstrating cytokeratin expression |
| Disclaimer |
1. The CoA for this product (provided upon request) verifies the cell-type specific gene expression via RT-PCR only. All test parameters provided in the CoA are conducted using abm's standardized culture system and procedures. The stated values may vary under the end-user's culture conditions. Please verify that the product is suitable for your studies by referencing published papers or ordering RNA (0.5 μg, Cat.# C207, $450.00) or cell lysate (100 μg, Cat.# C206, $600.00) to perform preliminary experiments, or alternatively use our Gene Expression Assay Service (Cat# C138). All sales are final.
2. We strongly recommend live cell shipments for ease of cell transfer and this option can be requested at the time of ordering. Please note that the end-user will need to evaluate the feasibility of live cell shipment by taking into account the final destination's temperature variation and its geographical location. In addition, we thoroughly test our cell lines for freeze-thaw recovery. If frozen cells were received and not recovered in your lab under the exact, specified conditions (using recommended culture vessel, media, additional supplements, and atmospheric conditions), a live cell replacement is possible at a cost (plus shipping).
3. All of abm's cell biology products are for research use ONLY and NOT for therapeutic/diagnostic applications. abm is not liable for any repercussions arising from the use of its cell biology product(s) in therapeutic/diagnostic application(s). Please contact a technical service representative for more information.
4. abm makes no warranties or representations as to the accuracy of the information on this site. Citations from literature and provided for informational purposes only. abm does not warrant that such information has been shown to be accurate. |
