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Immortalized-Human-Esophageal-Epithelial-Cells-(NE2-hTERT)

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Immortalized-Human-Esophageal-Epithelial-Cells-(NE2-hTERT)

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Immortalized-Human-Esophageal-Epithelial-Cells-(NE2-hTERT)

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	Print Version
BioSafety Level	II
Organism	Homo sapiens
Source Organ	Esophageal mucosa
Donor Age	52
Donor Gender	Male
Growth Properties	Adherent
Morphology	Epithelial-like
Recommended Seeding Density	Thaw entire contents into an appropriate T25 flask as specified in the Propagation instructions.
Markers	CK5, CK6, CK8, CK13, Ck18, CK19
Applications	For Research Use Only
Immortalization Method	Serial passaging and transduction with retroviruses carrying hTERT gene
Description	The Immortalized Human Esophageal Epithelial Cells (NE2-hTERT) retains epithelial morphology and can be used as a reliable supply of normal esophageal epithelial cells. The cells expresses cytokeratins 5, 6, 8, 13, 18, and 19. Whereas 6 and 18 are markers for proliferating epithelial cells while 8, 13, 18, and 19 are markers for esophageal squamous cells respectively. NE2-hTERT cells do not grow anchorage-independently and are not tumorigenic. It is recommended for applications in studies of esophageal epithelial cell functions and development of esophageal cancer.
Procedure Overview
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Propagation	The complete media for this cell line is 1:1 ratio of Defined Keratinocyte-SFM (DKSFM) with supplements (Gibco, #10744-019) and Epilife medium with EDGS (Cascade Biologics #M-EPI-500-CA; #S-012-5) . Atmosphere: air: 95%, CO₂: 5%; Temperature: 37.0°C.
Subculturing	To subculture from T-25 (1:3): 1. Remove media and wash with 2 m 1X PBS 2. Add 0.5 ml Trypsin-EDTA (TM050) (abm TM050) 3. Put flask into 37°C incubator for ~2 minutes or until cells detach 4. Neutralize using 3 ml of DKSFM supplemented with 2% FBS 5.Transfer cells to 15 ml tube and centrifuge at 700 rpm for 4 minutes at room temperature 6. Remove supernatant 7. Add 2 ml complete media to each flask 8. Resuspend cells with 1.5 ml complete medium 9. Add 0.5 ml resuspension into each flask containing 2 ml complete medium 10. Incubate in 37°C incubator with 5% CO₂ overnight 11. Next day remove media, and add 5 ml of complete media to each flask Be sure to use T25 ECM-coated flasks(G299) for growth.
Preservation	To freeze down from T-25 (1:2): 1. Remove media and wash with 2 m 1X PBS 2. Add 0.5 ml Trypsin-EDTA (abm TM050) 3. Put flask into 37°C incubator for ~2 minutes or until cells detach 4. Neutralize using 3 ml of DKSFM supplemented with 2% FBS 5. Transfer cells to 15 ml tube and centrifuge at 700 rpm for 4 minutes at room temperature 6. Remove supernatant 7. Add 2 ml complete media to each flask 8. Resuspend cells with 1 ml complete medium 9. Add 1ml of freezing medium (TM024) 10. Transfer 1 ml aliquot into each cryopreservation vial, and freeze 11. Transfer to liquid nitrogen tank the next day for long term storage
Quality Control	1) Immunostaining and Western blot were preformed for cell characterization
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.