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Immortalized-Drosophila-Embryonic-Cells-(R4)

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Immortalized-Drosophila-Embryonic-Cells-(R4)

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Immortalized-Drosophila-Embryonic-Cells-(R4)

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	Print Version
BioSafety Level	II
Organism	Drosophila
Source Organ	Embryos
Growth Properties	Adherent
Morphology	Spindle-shaped
Recommended Seeding Density	Thaw entire contents into an appropriate T25 flask as specified in the Propagation instructions.
Applications	For Research Use Only
Immortalization Method	Serial passaging of primary cultures from Act5C > UAS-RasV12 embryos
Description	The immortalized Drosophila Embryonic Cells (R4) was established by serial passaging of primary cultures from Act5C > UAS-Ras^V12 embryos. The Ras^V12 immortalized cell lines have significantly up-regulated cell-cycle and cell-division gene expression including gene targets of the E2 promoter binding factor/retinoblastoma protein (E2F/RB) pathway. Specifically, R4 express upregulated Cyc A, stg, dap, cdc2c, Cyc E, tum, sti, pav, stg, dup, dap, Orc2, and Mcm2. The cell lines are in a proliferative undifferentiated progenitor-like state associated with increasing levels of Polycomb Group (PcG) transcripts during the immortalization process. Related to *** muscle precursors (AMPs), a stem cell-like population contributing to *** muscles and sharing properties with vertebrate satellite cells, the immortalized R4 cell line retained the capacity for myogenic differentiation when treated with the steroid hormone ecdysone. The cell line is recommended as a tool for Drosophila embryogenesis and muscle cell differentiation and proliferation.
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 Schneider's Insect Medium (TM034). To make the completed growth medium, add the following components to the base medium at a final concentration of 10% heat-inactivated fetal bovine serum (TM999), and 1% Penicillin/Streptomycin (G255). Atmosphere: air: 95%, CO?: 5%; Temperature: 22°C.
Quality Control	(1) Inducing muscle cell differentiation by treating cells with ecdysone and perform immunofluorescence staining and qPCR to assess the upregulated expression of mhc and Tropomyosin (Tm). (2) Up-regulated genes in DPCs are analyzed by DAVID (the Database for Annotation, Visualization and Integrated Discovery).
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.