Don't look: growing clonal versus nonclonal neural stem cell colonies.
Stem Cells
; 26(11): 2938-44, 2008 Nov.
Article
em En
| MEDLINE
| ID: mdl-18757294
ABSTRACT
Recent reports have challenged the clonality of the neurosphere assay in assessing neural stem cell (NSC) numbers quantitatively. We tested the clonality of the neurosphere assay by culturing mixtures of differently labeled neural cells, watching single neural cells proliferate using video microscopy, and encapsulating single NSCs and their progeny. The neurosphere assay gave rise to clonal colonies when using primary cells plated at 10 cells/microl or less; however, when using passaged NSCs, the spheres were clonal only if plated at 1 cell/microl. Most important, moving the plates during the growth phase (to look at cultures microscopically) greatly increased the incidence of nonclonal colonies. To ensure clonal sphere formation and investigate nonautonomous effects on clonal sphere formation frequencies, single NSCs were encapsulated in agarose and proliferated as clonal free-floating spheres. We demonstrate that clonal neurospheres can be grown by avoiding movement-induced aggregation, by single-cell tracking, and by encapsulation of single cells. Disclosure of potential conflicts of interest is found at the end of this article.
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Células-Tronco
/
Esferoides Celulares
/
Neurônios
Limite:
Animals
Idioma:
En
Revista:
Stem Cells
Ano de publicação:
2008
Tipo de documento:
Article
País de afiliação:
Canadá