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$Immortalized-Mouse-Wildtype-M4<sup>+\/+<\/sup>-Epidermal-Keratinocyte-Cells$

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Immortalized-Mouse-Wildtype-M4^{+\/+<\/sup>-Epidermal-Keratinocyte-Cells}

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Immortalized-Mouse-Wildtype-M4^{+\/+<\/sup>-Epidermal-Keratinocyte-Cells}

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	+/+-Epidermal-Keratinocyte-Cells-T0459-Data-Sheet.html" target="_blank" rel="nofollow">Print Version
BioSafety Level	II
Organism	CD-1 Mouse
Source Organ	Skin
Growth Properties	Adherent
Morphology	Cobblestone-shaped
Recommended Seeding Density	Thaw entire contents into an appropriate T25 flask as specified in the Propagation instructions.
Markers	Keratin 14, Keratin 6
Applications	For Research Use Only
Immortalization Method	Immortalization in subsequent cultures
Description	The Immortalized Mouse Wildtype M4^+/+ Epidermal Keratinocyte Cells were derived from primary keratinocytes of newborn CD-1 mice. The primary keratinocytes were immortalized in subsequent cultures after passaging for an extended period. The immortalized cells display similar phenotype to the primary keratinocytes including the cobblestone-shape morphology. The Immortalized Mouse Wildtype M4^+/+ Epidermal Keratinocyte Cells are non-tumorigenic. They also differentiate with increased calcium, maintain a relatively stable karyotype no more aberrant than cultured primary keratinocytes, and produce epidermis upon orthotopic grafting onto nude mouse hosts. Upon introduction of v-ras^Ha oncogene, the Immortalized Mouse Wildtype M4^+/+ Epidermal Keratinocyte Cells produce squamous cell carcinoma in orthotopic skin grafts. These cells are useful in studies in epidermal keratinocyte biology and pathology.
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 VII medium available from abm (TM017). To make the completed growth medium, add the following components to the base medium: Fetal Bovine Serum (TM999) to a final concentration of 8%, 0.05mM calcium, 1ng/ml KGF, and 1% Penicillin/Streptomycin (G255). Atmosphere: air: 95%, CO?: 5%; Temperature: 37.0°C.
Subculturing	1. Remove and discard culture medium. 2. Add 2.0mL of Trypsin-EDTA (TM050) solution to flask and observe cells under an inverted microscope until cell layer is dispersed. Note: Cells that are difficult to detach may be placed at 37°C to facilitate dispersal. 3. Centrifuge cells at 1500rpm for 3 minutes to pellet. 4. Aspirate out trypsin, leaving pellet undisturbed. 5. Resuspend pellet in fresh culture medium and plate in new culture vessel. 6. Incubate cultures at 37°C. Be sure to use T25 ECM-coated flasks(G299) for growth.
Preservation	1. Freeze Medium: Complete growth medium with 20% FBS and 10% DMSO. 2. Storage Temperature: Liquid nitrogen vapour phase.
Quality Control	1) Immunofluorescence analysis of Keratin markers
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.