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$Immortalized-Human-Cervical-Keratinocytes-(16E6\/E7-HCK)$

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Immortalized-Human-Cervical-Keratinocytes-(16E6\/E7-HCK)

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Immortalized-Human-Cervical-Keratinocytes-(16E6\/E7-HCK)

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	Print Version
BioSafety Level	II
Organism	Homo sapiens
Source Organ	Cervix
Growth Properties	Adherent
Morphology	Polygonal
Recommended Seeding Density	Thaw entire contents into an appropriate T25 flask as specified in the Propagation instructions.
Applications	For Research Use Only
Immortalization Method	Transfection with retrovirus expressing HPV-16 E7 and HPV-16 E6
Description	The Immortalized Human Cervical Keratinocytes (16E6/E7 HCK) were derived from normal human ectocervix cells from surgical specimens and were immortalized by transfection with retrovirus expressing HPV-16 E7 and E6. These immortalized cells are suitable as a model for carcinogenesis and cellular senescence as the expression of HPV-E6 and E7 proteins, and inactivation of both p53 and Rb, extend the lifespan of cells past a first critical period when the normal cells senesce. Having telomere restoration ability, the Immortalized Human Cervical Keratinocytes (16E6/E7 HCK) are also useful for studies focusing on understanding the role of telomere loss and restoration in the development of HPV-associated human cancers.
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 growth medium for this cell line is Keratinocyte-SFM (Gibco-BRL, Gaithersburg, MD) with 1% Penicillin/Streptomycin Solution available from abm (G255). Atmosphere: air: 95%, CO?: 5%; Temperature: 37.0°C.
Quality Control	1) FACS with propidium iodide staining; 2) RIPs to detect HPV-16 E6 and E7.
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.