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Immortalized-Mouse-Lung-Epithelial-Cells-(MLE-15)

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Immortalized-Mouse-Lung-Epithelial-Cells-(MLE-15)

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Immortalized-Mouse-Lung-Epithelial-Cells-(MLE-15)

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	Print Version
BioSafety Level	II
Organism	SP-C/TAg Transgenic Mice
Source Organ	Distal respiratory epithelium
Donor Gender	Female
Growth Properties	Adherent
Morphology	Polygonal
Recommended Seeding Density	Thaw entire contents into an appropriate T25 flask as specified in the Propagation instructions.
Markers	SP-A (low level), SP-B, SP-C
Applications	For Research Use Only
Description	Nonciliated bronchiolar (Clara) cells, alveolar type I and type II cells are the predominant cell types in the respiratory epithelium. Clara cells and alveolar type II cells produce pulmonary surfactant proteins that are critical in the structure and function maintenance of the pulmonary alveolus by reducing surface tension at the air-liquid interface. The Immortalized Mouse Lung Epithelial Cells (MLE-15) is a cell line derived from transgenic FVB/N mice harbouring SV40 TAg under the control of the human SP-C promoter. Similar to alveolar type II cells, MLE-15 is characterized by the presence of apical microvilli and intracellular multiamellar inclusion bodies, as well as the ability to produce surfactant proteins B and C. This cell line is useful in studying the regulation of lung epithelial genes and elucidating the complex transcriptional networks related to airway/alveolar differentiation.
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 II medium available from ABM (TM002). To make the complete growth medium, add the following components to the base medium: 2% fetal bovine serum (TM999), 5 mg/L insulin, 10 μg/ml transferrin, 3 ng/ml sodium selenite, 10 nM hydrocortisone, 10 nM β-estradiol 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) Electron microscopy was used to evaluate the presence of microvilli and lamellar bodies; 2) in situ hybridization was performed to assess the presence of respiratory epithelial cell markers; 3) Immunoprecipitation was performed to determine the secretion of surfactant proteins; 4) Northern blot was used to detect SP-A, SP-B and SP-C, and SV40 TAg mRNA.
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.