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Isolation and manipulation of mammalian neural stem cells in vitro.

Methods in molecular biology (Clifton, N.J.) (2008-12-18)
Claudio Giachino, Onur Basak, Verdon Taylor
ABSTRACT

Neural stem cells are potentially a source of cells not only for replacement therapy but also as drug vectors, bringing bioactive molecules into the brain. Stem cell-like cells can be isolated readily from the human brain, thus, it is important to find culture systems that enable expansion in a multipotent state to generate cells that are of potential use for therapy. Currently, two systems have been described for the maintenance and expansion of multipotent progenitors, an adhesive substrate bound and the neurosphere culture. Both systems have pros and cons, but the neurosphere may be able to simulate the three-dimensional environment of the niche in which the cells reside in vivo. Thus, the neurosphere, when used and cultured appropriately, can expand and provide important information about the mechanisms that potentially control neural stem cells in vivo.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Trypsin inhibitor from Glycine max (soybean), powder, BioReagent, suitable for cell culture
Sigma-Aldrich
Bovine Serum Albumin, heat shock fraction, protease free, pH 7, ≥98%
Sigma-Aldrich
L-Cysteine hydrochloride, anhydrous, from non-animal source, BioReagent, suitable for cell culture, ≥98.0%
Sigma-Aldrich
Papain from papaya latex, buffered aqueous suspension, 2× Crystallized, ≥16 units/mg protein