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Immortalized-Rat-Sympathoadrenal-Progenitor-Cells-(MAH-A3)

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Immortalized-Rat-Sympathoadrenal-Progenitor-Cells-(MAH-A3)

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Immortalized-Rat-Sympathoadrenal-Progenitor-Cells-(MAH-A3)

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	Print Version
BioSafety Level	II
Organism	E14.5 Rat Embryo
Source Organ	Adrenal Glands
Growth Properties	Adherent
Morphology	Round
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 and transduction with recombinant lentiviruses carrying v-myc oncogene
Description	The Immortalized Rat Sympathoadrenal Progenitor Cells (MAH-A3) are derived from HNK-1+ cell population isolated from embryonic rats and that are immortalized using the v-myc oncogene. The cells exhibit morphology resembling the primary cells, and expresses progenitor-specific markers (HNK-1, TH, SCG10, and NF68) and surface antigens such as L1, N-CAM, and Thy-1. MAH-A3 cells are undifferentiated precursors that do not express NGF receptors; however but when cultured in the presence of basic fibroblast growth factor (bFGF abm Z101455), the cells become NGFR-positive post-mitotic sympathetic neurons. MAH-A3 cells are research tools recommended to study the sympathetic development mechanisms.
Procedure Overview
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Propagation	Grow cells in T25 poly-D-lysine and laminin-coated flasks with the following conditions. The base medium for this cell line is L15-CO2 medium (Lonza; 12-669E). To make the complete growth medium, add the following components to the base medium: fetal bovine serum (TM999) to final concentration of 10%, 50 μM dexamethasone, and Penicillin/Streptomycin (G255). Temperature: 37.0°C. To differentiate cells: The base medium for this cell line is L15-CO2 medium. To make the complete growth medium, add the following components to the base medium: fetal bovine serum (TM999) to final concentration of 10%, 10 ng/ml bFGF(Z101455), and Penicillin/Streptomycin (G255). Note: FGF will increase cell proliferation and neurite outgrowth, but the removal of dexamethasone causes the cells to die more rapidly. It is recommended to grow cells in the presence of dexamethasone before culturing cells to cause differentiation.
Quality Control	1) Northern blot to confirm v-myc-containing pro-viral RNA transcripts and cell markers; 2) Southern blot to analyze pro-viral transcript integration; 3) Immunofluorescence staining of sympathoadrenal 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.