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Immortalized-Human-Corneal-Endothelial-Cells-(IHCE)

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Immortalized-Human-Corneal-Endothelial-Cells-(IHCE)

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Immortalized-Human-Corneal-Endothelial-Cells-(IHCE)

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	Print Version
BioSafety Level	II
Organism	Homo sapiens
Source Organ	Descemets′ membrane
Donor Gender	Male
Growth Properties	Adherent
Morphology	Cobblestone-like
Population Doubling	54 hours
Recommended Seeding Density	20, 000 cells/cm²
Markers	CX43, ZO-1, 9.3E
Applications	For Research Use Only
Immortalization Method	Serial passaging and transduction with retroviruses carrying human papillomavirus type 16 E6/E7 gene
Description	The role of the corneal endothelium is to regulate corneal hydration, and dysfunction of this critical cellular layer will gradually result in corneal opacification and eventually results in loss of vision and corneal blindness. Established using the HPV16 E6/E7 genes, The Immortalized Human Corneal Endothelial Cells (IHCE) not only retains its authentic cell biomarkers (CX43, ZO-1 and 9.3E), it also displays fairly disomic cytogenetic status at chromosomes 1, 8, 10 and 18, which is an indication of normal diploid chromosomal status. Together with the Immortalized Human Corneal Keratinocytes (T0579) and the Immortalized Human Corneal Fibroblasts (T0578) isolated from the same donor and immortalized using the same platform, this uniform model system presents a valuable tool for studies involving cornea homeostasis and ophthalmologic diseases.
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 I (TM001). To make the complete growth medium, add the following components to the base medium: 10% fetal bovine serum (TM999), 5 mg/L human insulin (Z101065), 10 μg/ml transferrin, 3 ng/ml sodium selenite, 10 nM hydrocortisone, 10 nM β-estradiol, 10ng.ml VEGF, 10ng/ml hEGF, 10ng/ml Heparin, 1% L-glutamine and 1% Penicillin/streptomycin (G255). Atmosphere: air: 95%, CO2: 5%; Temperature: 37.0°C.
Preservation	1. Freeze Medium: Complete growth medium with 20% FBS and 10% DMSO. 2. Storage Temperature: Liquid nitrogen vapour phase.
Quality Control	1) RT-PCR was used to evaluate the transgene expression of HPV16 E6/E7; 2) Indirect immunofluorescence was performed to detect the cell specific marker expression CX43, ZO-1 and 9.3E; 3) FISH was used to examine the cytogenetic alterations in the immortalized cells.
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.