It was recently reported that the SCZ-aNSCs have therapeutic potential in traumatic brain injury 9. The subcallosal zone (SCZ) of the caudal subcortical white matter is recognized as a novel neurogenic region 6-8. NSCs reside in different brain regions, such as the subventricular zone (SVZ) of the lateral ventricle and the subgranular zone (SGZ) of the hippocampus 3-5. These two well-developed culture systems have contributed to the characterization of aNSCs in vitro. Compared to the neurosphere culture, the monolayer culture system provides better homogenous maintenance and expansion of NSCs 2. Monolayer cultures are also widely used for the maintenance and differentiation of NSCs. The number of cells and the size of the neurosphere are considered to be closely associated with the proliferation properties of the NSCs. Depending on self-renewal potency, NSCs continuously proliferate and generate a cell mass in a suspension culture. The neurosphere culture system was developed in the early 1990s and served as a standard stem cell culture system 1. To confirm these properties, a neurosphere culture system has widely been used. NSCs have characteristics of self-renewal and multiple-lineage differentiation. Thus, using a discrete brain region, these culture systems will be helpful for expanding our knowledge about aNSCs and their applications for therapeutic uses. Significantly, this culture system provides a more homogenous environment for NSCs than the neurosphere culture system. A monolayer culture is also an in vitro system to assay proliferation and differentiation. A neurosphere culture system provides a useful tool for determining proliferation and collecting the genuine NSCs.
#ADULT NEURAL STEM CELLS VERIFICATION#
A well-developed culture system allows for the verification of their characteristics and can increase research on NSCs. The SCZ sections are initially subjected to a neurosphere culture. A brain matrix with intervals of 1 mm is used to obtain the SCZ-containing coronal slices and to precisely dissect the SCZ from the whole brain. This protocol describes in detail the process to generate SCZ-aNSCs from an adult mouse brain. These cells are suggested to have a therapeutic potential for traumatic cortical injury. A low percentage of aNSCs from the SCZ is differentiated into neurons most cells are differentiated into glial cells, such as oligodendrocytes and astrocytes. The subcallosal zone (SCZ), located between white matter and the hippocampus, has recently been reported to contain aNSCs and continuously give rise to neuroblasts. Identifying aNSC-derived regions and characterizing the aNSC properties are critical for the potential use of aNSCs and for the elucidation of their role in neural regeneration. NSCs have the potential for differentiation and proliferation into three types of cells: neurons, astrocytes, and oligodendrocytes. Adult neural stem cells (aNSCs) can be used for the regeneration of damaged brain tissue.
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