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$Immortalized-Mouse-Retinal-Pigmented-Epithelial-Cells---HPV16-E6\/E7$

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Immortalized-Mouse-Retinal-Pigmented-Epithelial-Cells---HPV16-E6\/E7

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Immortalized-Mouse-Retinal-Pigmented-Epithelial-Cells---HPV16-E6\/E7

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	Print Version
BioSafety Level	II
Organism	Mus musculus C57B1/6
Source Organ	Retina
Growth Properties	Adherent
Morphology	Cobblestone-like
Recommended Seeding Density	Thaw entire contents into an appropriate T25 flask as specified in the Propagation instructions.
Markers	1. RPE65 and CRALBP (Differentiated RPE) 2. ZO1, Cytokeratin -8 and -18 (Epithelial cell marker)
Applications	For Research Use Only
Immortalization Method	Serial passaging and transduction with retrovirus carrying human papiloma virus 16 E6/E7
Description	The Immortalized Mouse Retinal Pigmented Epithelial Cells- HPV E6/E7, derived from healthy C57B1/6 mouse RPE cells, is a cell line that retained many of their in vivo phenotypic characteristics. RPE-specific markers such as RPE65 and cellular retinaldehyde binding protein (CRALBP), as well as epithelial markers ZO1, cytokeratin 8 and 18 are expressed in the Immortalized Mouse Retinal Pigmented Epithelial Cells- HPV E6/E7. These cells are useful in studies of RPE cell biology and investigation of ocular disease including age related macular degeneration (AMD).
Procedure Overview
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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 III medium available from ABM (TM003). To make the complete growth medium, add the following components to the base medium: fetal bovine serum (TM999) to a final concentration of 10%, Penicillin/Streptomycin(G255). Atmosphere: air: 95%, CO2: 5%; Temperature: 37.0°C.
Quality Control	1. Western Blot was used to verify the HPV protein expression in the immortalized cell line. 2. Western blot and real-time RT-PCR was used to confirm the presence of in vivo RPE markers such as RPE65 and CRALBP. 3. Immunofluorescence staining was used to confirm expression of epithelial cell markers ZO1, cytokeratin 8 and cytokeratin 18. 4. Absence of neural cell marker, SV2, PKCα, rhodopsin
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.