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Immortalized-Human-Cystic-Fibrosis-Tracheobronchial-Epithelial-Cells-(CFT1)

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Immortalized-Human-Cystic-Fibrosis-Tracheobronchial-Epithelial-Cells-(CFT1)

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Immortalized-Human-Cystic-Fibrosis-Tracheobronchial-Epithelial-Cells-(CFT1)

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	Print Version
Biosesafety Level	II
Organism	Homo sapiens
Source Organ	Trachea
Donor Age	24 y/o
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.
Applications	For Research Use Only
Immortalization Method	Transfection with the pUC19 plasmid encoding intact HPV-18 E6 and E7 gene
Description	Cystic fibrosis (CF) is a genetic disease affecting the exocrine glands in which production of abnormal thick mucus causing build up in the bronchi, pancreatic ducts, and the intestines which may lead to blockages and eventually resulting in respiratory infection. The Immortalized Human Cystic Fibrosis Tracheobronchial Epithelial Cells (CFT1) is an applicable in vitro model for further research in CF which exhibits differentiated phenotypes in ion transport and have a decrease regulation of Cl- ion channel. The primary cells were derived from a 24 year-old, male donor which the history of meconium ileus at birth, pancreatic insufficiency, elevated sweat Cl-, bronchiectatic lung disease with chronic Staphylococcus aureus and Pseudomonas aeruginosa infection, and was determined to be homozygous for phenylalanine-508 deletion in the CF transmembrane conductance regulator (CFTR) gene. Immortalization with HPV-18 E6/E7 established a continuous cell line for experimental studies for the CFTR gene and function.
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 IV medium available from abm (TM004). To make the completed growth medium, add the following components to the base medium in the specified final concentrations: 10 µg/ml insulin (), 10^-6M hydrocortisone, 3.75 µg/ml endothelial cell growth supplement (Sigma; E2759), 25 ng/ml EGF (), 3x10^-8M T3 (triiodothyronine) (Sigma; T06397), 5 µg/ml Transferrin (Sigma; T0665), 10 ng/mlCholera toxin (Sigma; C-3012), and Penicillin/Streptomycin (G255). Atmosphere: air: 95%, CO₂: 5%; Temperature: 37.0°C.
Quality Control	Immunoblot used to determine the expression of CFTR, and presence of HPV-18 E6/E7 gene.
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 evalsuate the feasibility of live cell shipment by taking into account the final destination's temperature variation and its geographical locations. 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.