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Mouse Primary Neural Stem Cells

SKU : BHC10901851

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Applied Biological Materials (abm) Inc.

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Mfr.No.	T4173
Cell Type	Primary Cells
Biosafety Level	2
Disclaimer	1. Sale of this item is subjected to the completion of a Material Transfer Agreement (MTA) by the purchasing individual/institution for each order. If you have any questions regarding this, please contact us at [email protected]. 2. 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 ?g, Cat.# C207, $450.00) or cell lysate (100 ?g, Cat.# C206, $600.00) to perform preliminary experiments, or alternatively use our Gene Expression Assay Service (Cat# C138). All sales are final. 3. We 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). 4. 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. 5. 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. 6. abm warrants that cell lines shall be viable upon initiation of culture for a period of thirty (30) days after shipment and that they shall meet the specifications on the applicable abm Material Product Information sheet, certificate of analysis, and/or catalog description. Such thirty (30) day period is referred to herein as the "Warranty Period."
Disease	Normal
Donor History	Normal tissue
Growth Properties	Suspension, neural sphere
Growth Conditions	Seed cells using 10 cm petri dishes. PriGrow X Series Medium (TM4173) + 1% Penicillin/Streptomycin Solution (G255), 37.0?, 5% CO?.Neurosphere Dissociationg Solution for T4173 (TM4173-DS) is required for dissociating neurospheres.
Quantity	5x10⁵ cells / 1.0 ml
Tissue	Brain
Morphology	Neural Sphere
Notes	1. Visually examine the packaging containers for signs of leakage or breakage. 2. Immediately transfer frozen cells from dry ice packaging to a temperature below -130?, preferably in liquid nitrogen vapor phase storage, until ready for use.To ensure the highest level of viability, thaw the vial and initiate culture as soon as possible upon receipt. If continued storage is desired, the vial should only be stored below -130? or in liquid nitrogen vapor phase. Do not store at -70?, as it will result in loss of viability.
Organism	Mouse (M. musculus)
Species	Mouse
Propagation	PriGrow X Series Medium (TM4173) + 1% Penicillin/Streptomycin Solution (G255), 37.0°C, 5% CO?.
Cryopreservation	Cryopreservation Medium (TM024), or complete growth media with 10% DMSO.
Source Organ	Brain

Applications Range	Research Use Only.

Shipping	Ship with dry ice.
Product Format	Frozen
Product Use	This product is intended for laboratory research use only. It is not intended for any animal or human therapeutic use, any human or animal consumption, or any diagnostic use.
Storage	Vapor phase of liquid nitrogen, or below -130?.

Thawing Protocol	1. Thaw cells quickly in a 37? water bath while agitating gently (maximum 2 minutes). The vial cap should be kept above the water level to minimize the risk of contamination. 2. Decontaminate the vial by spraying and wiping the exterior of the vial with 70% ethanol. From this point onwards, all operations should be strictly carried out inside a biological safety cabinet using aseptic conditions. 3. Transfer the cell suspension into a 15ml sterile conical tube containing 9ml of pre-warmed, complete growth media. Centrifuge cells at 200xg for 5 minutes. 4. Aspirate the supernatant without disturbing the cell pellet. Re-suspend the cell pellet in the recommended pre-warmed, complete growth media and dispense into a T25 culture flask. Cells should float without attaching.?br />5. Incubate the cells at the recommended conditions. 6. Cells will form floating neurospheres in 2-3 days. Do not let neurospheres attach to the cutlure vessel; gently pipette area to loosen the attached neurospheres, as frequently as required.?br />7. Perform hemi-depletions every 2-3 days by tilting the culture vessel and allowing the neurospheres to settle in one corner of the flask. Remove half of the medium and replace with fresh medium without disturbing the floating cells.?br />8. Dissociate neurosphere when they reach 100-200?m in diameter.
Subculture Protocol	To disscoiate neurospheres: 1. Warm the Neurosphere Dissociation Solution (TM4173-DS) to room temperature.?br>2. Collect neurospheres in a 15ml sterile conical tube?nd centrifuge at 800rpm for 3-5 minutes.?br>3. Aspirate the supernatant without disturbing the cell pellet and then gently flick the tube to loosen the neurosphere pellet.?br>4. Resuspend neurospheres in 3-5ml of the Neurosphere Dissociating Solution and incubate in a 37C water bath for 2 minutes.?br>5. Add 10 ml of complete growth medium and spin cells at 1200 rpm for 5 minutes.?br>6. Carefully aspirate the supernatant and resuspend the pellet in 1 ml of complete growth medium.?br>7. Gently?riturate 10-15 times with a fire polished glass pipette to break down spheres.?br>8. Allow the undigested clusters to settle to the bottom and transfer the cells in suspension to a new tube.?br>9. Add additional media to the undigest clusters and repear step 7 and combines with the suspension cells collected previously.?br>10. Plate cells as recommended.

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SKU	Supplier No.	Size	Your price	Quantity
BHC10901851	T4173	5x10^5 cells / 1.0 ml	-		Add to Quote

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