Application protocol

Isolation and cultivation of adult 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 the dissociation, the cell debris were removed using the Debris Removal Solution. NSCs were stained using the Adult Neural Stem Cell Sorting and Analysis Kit, 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.

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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Protocol


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