Protocol abstract
Nature Protocols 3, - 935 - 940 (2008)
Published online: 8 May 2008 | doi:10.1038/nprot.2008.55
Subject Categories: Cell and tissue culture | Isolation, purification and separation | Model organisms
Production of neurons, astrocytes and oligodendrocytes from mammalian CNS stem cells
Andrew Chojnacki1 & Samuel Weiss1
Abstract
The isolation and expansion of precursor cells in a serum-free culture system allows for the systematic characterization of their properties and the intrinsic and extrinsic signals that regulate their function. The discovery of neural stem cells in the adult mouse brain was made possible by the creation of a novel culture system subsequently termed the neurosphere assay. Therein, the dissociated adult mouse periventricular area was plated in the presence of epidermal growth factor, but in the absence of adhesive substrates, which resulted in the generation of spheres of proliferating cells that detached from the plate bottom and remained suspended in the media. Since its inception, the neurosphere culture system has been widely used in the neural precursor cell field and has been extensively adapted for the isolation and expansion of corneal, cardiac, skin, prostate, mammary and brain tumor stem cells. The original neurosphere culture protocol, which takes approximately 10 d to complete, is described here in detail.
- Hotchkiss Brain Institute, Department of Cell Biology and Anatomy, Faculty of Medicine, University of Calgary, Calgary, Alberta Canada, T2N 4N1.
Correspondence to: Samuel Weiss1 e-mail: weiss@ucalgary.ca
nature-products
A-Z product listing
- 0.1-mm-deep hemacytometer(Hausser Scientific)
- 0.22-
m bottle-top filter(Millipore) - 10
Dulbecco's modified Eagle's media(Invitrogen/Gibco) - 10 Dulbecco's PBS(Invitrogen/Gibco)
- 10-cm Petri dish(Fisher)
- 14-ml polypropylene round-bottomed tubes(BD Falcon)