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Immortalized-Mouse-Neuronal-Cells-(M4b)

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Immortalized-Mouse-Neuronal-Cells-(M4b)

商品詳情參考文獻(xiàn) 相關(guān)資料

	Print Version
BioSafety Level	II
Organism	Mouse
Source Organ	Spinal Cord
Growth Properties	Adherent
Morphology	Neuronal
Population Doubling	14 hours
Recommended Seeding Density	40,000 cells/cm²
Applications	For Research Use Only
Immortalization Method	Supplementation of plating medium with UCHT1 rat thyroid cell line-conditioned medium
Description	The Immortalized Mouse Neuronal Cells (M4b) were derived from the spinal cord of a fetal mouse after 12-16 days of gestation. The fetal mouse was bred from double heterozygous (Rb 2H/RB 32 Lub) males and normal C57BL females. Transformation of the cell line was induced via supplementation with UCHT1 rat thyroid cell line-conditioned medium and was confirmed via the detection of vimentin. Immunohistochemistry found the presence of neurological makers, but the absence of glial markers in the Immortalized Mouse Neuronal Cells (M4b). In addition, the intracellular [Ca²⁺] response after neurotransmitter stimulation was rapid and transient as expected. The Immortalized Moue Neuronal Cells (M4b) is useful in studies of neuronal cell biology, viral infections, and neuronal cytotoxicity. The Immortalized Moue Neuronal Cells (M4b) can also be used as a control for the Immortalized Trisomy 16 Mouse Neuronal Cells (MTh) (Cat. T0675), which is a mouse model of Down Syndrome.
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: Fetal Bovine Serum (TM999) to final concentration of 10%, Bovine Serum (Sigma) to a final concentration of 10%, 1% L-glutamine (G275) 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) RT-PCR for SOD-1 and GAPDH; 2) Immunohistochemistry of neuronal markers; 3) Intracellular Ca²⁺ measurements;
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.