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Immortalized-Human-Hepatocytes---HPV-and-hTERT

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Immortalized-Human-Hepatocytes---HPV-and-hTERT

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Immortalized-Human-Hepatocytes---HPV-and-hTERT

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	Print Version
BioSafety Level	II
Organism	Homo sapiens
Source Organ	Liver
Growth Properties	Adherent
Passage Number	>30
Population Doubling	40-60 hours
Recommended Seeding Density	4 x 10⁴cells/cm²
Applications	For Research Use Only
Immortalization Method	Serial passaging and transduction with lentiviruses carrying Human papillomavirus E6/E7 protein and hTERT gene.
Description	Human hepatocytes plays an important role in life science research for evaluation of key human-specific drug properties, such as of drugs interactions, toxicity tests and metabolism reactions. Since primary cells are often hard to propagate in culture abm offers immortalized human hepatocytes that allows the cells to escape senescence crisis and help further research needs by extending the passage ability that primary cells lacks. These cells are ready-to-use for applications in research and developmental viral infections, screening, co-cultures and transfections.
Procedure Overview
Propagation	These cells requires collagen coated vessel, the collagen coating solution is available in abm TM101. The base medium for this cell line is Prigrow X series medium available in abm, Cat. No. TM0212. Atmosphere: air, 95%; Carbon dioxide (CO₂), 5%. Temperature: 37.0°C. We do not recommend the use of antibiotic for this cell line.
Subculturing	1. Remove and discard culture medium. 2. Add 2.0mL of Trysin-EDTA solution (TM102) to T25 flask and observe cells under an inverted microscope until cell layer is dispersed. Note: Continuously bump the vessel gently to facilitate dispersal of the cells. Once the cells are detached, add equal volume of complete medium to flask. 3. Centrifuge cells at 180 x g for 3 minutes to pellet. 4. Aspirate out supernatant, leaving pellet undisturbed. 5. Resuspend pellet in fresh culture medium and plate in new collagen coated culture vessel. 6. Incubate cultures at 37°C.
Preservation	1. Freeze Medium: Complete growth medium with 20% FBS and 10% DMSO. 2. Storage Temperature: Liquid nitrogen vapour phase.
Quality Control	Real Time PCR was used to quantify HPV E6/E7 and hTERT gene expression in immortalized cell line.
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.