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BioSafety Level	II
Organism	6- to 8-week old C57B/6 female mice
Source Organ	Mammary Epithelia
Growth Properties	Adherent
Morphology	Cuboidal
Passage Number	3
Recommended Seeding Density	Thaw entire contents into an appropriate T25 flask as specified in the Propagation instructions.
Markers	β-catenin, E-cadherin, Ep-CAM, CK8 and CK18
Applications	For Research Use Only
Immortalization Method	Serial passaging and electroporation transfection with linearized DNA of E1A and p53DD
Description	The Immortalized Mouse Mammary Epithelial Cells (iMMEC) is derived by co-expression of adenovirus E1A and dominant-negative p53 (p53DD). This cell line is unique in that 1) it expresses surface markers, including estrogen-receptor alpha (ERα) and retains the functional properties characteristic to mammary epithelial cell and 2) upon lactogenic stimulation it forms dome-like structures indicative of mammary alveoli and secrets β-catenin into the acinar lumen. This cell line is suitable for studies in hormone regulation and 3D dimensional ductal morphogenesis.
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: 10% fetal bovine serum (TM999), 5 μg/ml insulin, 1 μg/ml hydrocortisone and 5ng/ml EGF. Atmosphere: air: 95%, CO?: 5%; Temperature: 37.0°C. Lactogenic stimulation can be accomplished by inducing the cells with differentiation medium for 6 days (with media change every 2 days). To make complete differentiation medium, add the following components to the basal Prigorw IV medium (TM004): 5 μg/ml insulin, 1 μg/ml hydrocortisone, 3 μg/ml prolactin and 2% Matrigel.
Quality Control	1. Western blot was used to confirm the presence of E1A and p53DD gene in the immortalized cell line. 2. Western blot and immunofluorescence were used to check gene expression of β-catenin, E-cadherin, Ep-CAM, CK8, CK18 and ERα.
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.
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聯系方式	何經理，合肥星肽生物科技有限公司，電話：0551-63802898 400-8702-898，                                                  電郵：info@qingbio.com http://www.huangyuansheng.cn QQ：514713116

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