Application protocol

Isolation and cultivation of neural stem cells from adult mouse brain

 

This application protocol describes a step-by-step protocol to isolate pure neural stem cells (NSCs) from the subventricular zone (SVZ) of adult mouse brain. Brain tissue from adult mice was dissociated into single-cell suspensions using the Neural Tissue Dissociation Kit (T) in combination with the gentleMACS™ Dissociator with Heaters. After dissociation, cell debris was removed using the Debris Removal Solution. NSCs were stained using the Adult Neural Stem Cell Analysis Cocktail Kit, anti-mouse and sorted using the MACSQuant® Tyto® Cell Sorter. The MACSQuant Analyzer 10 was used for flow cytometric acquisition and data analysis. Neurosphere cultivation and differentiation assays were performed afterwards using the MACS® Neuro Medium and MACS NeuroBrew®-21 w/o Vitamin A.

Protocol

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This protocol includes detailed instructions for preparing necessary reagents and materials and then carrying out each step. 

Materials

The following is a listing of reagents, instruments, and consumables needed for each step of this protocol. These products are for research use only. 
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