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Immortalized-Mouse-Lung-Endothelial-Cells---PmT

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Immortalized-Mouse-Lung-Endothelial-Cells---PmT

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Immortalized-Mouse-Lung-Endothelial-Cells---PmT

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	Print Version
BioSafety Level	II
Organism	Mouse
Source Organ	Lung
Growth Properties	Adherent
Morphology	Endothelial
Recommended Seeding Density	Thaw entire contents into an appropriate T25 flask as specified in the Propagation instructions.
Applications	For Research Use Only
Immortalization Method	Immortalized by infecting with polyoma middle T (PmT) virus
Description	Primary mouse lung endothelial cells were derived and immortalized with polyoma middle T (PmT) virus to generate the Immortalized Mouse Lung Endothelial Cells – PmT. The cells are valuable for downstream applications to study the lung endothelium, as they can continuously divide in culture. It may be useful for studies relating to diseases and understanding the biology of lung endothelial cells.
Procedure Overview
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. This cell line requires coated cell culture vessels of collagen (final concentration of 30ug/ml; G422) and fibronectin (final concentration of 10ug/ml; TM059) in 0.1% gelatin (TM063). The base medium for this cell line is a 1:1 (recommend 400ml:400ml) ratio mix of PriGrow VI and PriGrow IV Series medium available in abm, Cat. No. TM016, and Cat. No. TM004 respectively. To make the completed growth medium, add the following components to the base medium: 100 mg heparin (Sigma; H3142) dissolved in 10 ml of PriGrow IV Medium (TM004), 10 ml Penicillin/Streptomycin (G255), and 20 ml L-glutamine (G275). Filter the medium with 0.2 uM filter before adding fetal bovine serum (TM999) which is first heat-inactivated before use to final concentration of 10% and 50 mg of endothelial mitogen (Generon). Atmosphere: air: 95%, CO?: 5%; Temperature: 37.0°C. Recommended split ratio of 1:4 or 1:8. Preform a PBS wash before using 0.25% Trypsin-EDTA (TM050) to subculture.
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.