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Immortalized-Mouse-Astrocytes---SV40T-(IMA2.1)

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Immortalized-Mouse-Astrocytes---SV40T-(IMA2.1)

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Immortalized-Mouse-Astrocytes---SV40T-(IMA2.1)

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	Print Version
Biosesafety Level	II
Organism	Wildtype BALB/c mice
Source Organ	Cerebral Cortex
Growth Properties	Adherent
Morphology	star-shape morphology
Recommended Seeding Density	Thaw entire contents into an appropriate T25 flask as specified in the Propagation instructions.
Markers	Nestin, CD44
Applications	For Research Use Only
Immortalization Method	Serial passaging and transfection with psV3neo plasmid carrying SV40 large T antigen genes
Description	The Immortalized Mouse Astrocytes- SV40 T (IMA 2.1) is a suitable cell line for studies in astrocyte functions. This cell line is unique in that it is capable of metabolizing the parkinsonian toxin MPTP (to MPP+) and is responsive to pro-inflammatory stimuli. In addition, this cell line can be used in neuronal co-culture studies to simulate conditions in the brain. Upon stimulation with cytokines such as IL-1β, TNF-α and IFN-γ, these cells show up-regulated NOS-2 and COX-2 mRNA expression and release various inflammatory mediators. This cell line has a stable star-shape morphology and expresses immature astrocyte markers such as nestin and CD44, indicating that the cells are of the early-stage astrocyte precursor phenotype. Furthermore, they are capable of converting glutamate via glutamine synthase to glutamine, which is a characteristic of GFAP-negative astrocytes.
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 III medium available from ABM (TM003). To make the complete growth medium, add the following components to the base medium: fetal bovine serum (TM999) to a final concentration of 10% and 1% Penicillin/Streptomycin (G255). Atmosphere: air: 95%, CO₂: 5%; Temperature: 37.0°C.
Quality Control	The cell line's response to pro-inflammatory stimuli was measured by 1) immunostaining to assess NF-kB translocations from the cytosol to the nucleus; 2) RT-qPCR and quantitative PCR detection of NOS-2 mRNA levels; and 3) Western Blot analysis of NOS-2 and COX2 proteins.
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 evalsuate the feasibility of live cell shipment by taking into account the final destination's temperature variation and its geographical locations. 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.