A non-contact suspension culture approach to the culture of osteogenic cells derived from a CD49elow subpopulation of human bone marrow-derived cells.
Biotechnol Bioeng
; 98(6): 1195-208, 2007 Dec 15.
Article
en En
| MEDLINE
| ID: mdl-17614333
We demonstrate that adult human bone marrow (BM) contains a population of mesenchymal stromal cells (MSCs) that can be expanded in non-adherent, cytokine-dependent, suspension culture conditions for at least 42 days. The cells generated during suspension culture lacked detectable levels of gene expression associated with differentiated mesenchymal cell types, including bone, muscle and fat, suggesting that suspension culture maintains MSCs in an uncommitted state. However, when these undifferentiated cells were taken out of suspension culture and placed in adherent osteogenic conditions, osteogenic genes were upregulated and morphologically identifiable bone matrix was elaborated. Flow cytometric analysis of uncultured, density gradient-separated human BM revealed that colony forming unit-fibroblast (CFU-F) and CFU-osteoblast (CFU-O) activity was associated with a CD45(-) CD49e(low) phenotype. Importantly, suspension-grown MSCs, capable of CFU-F and CFU-O development, maintained the CD45(-)CD49e(low) phenotype whereas MSCs directly cultured under adherent conditions rapidly upregulated CD49e expression and were associated with a CD45(-)CD49e(high) phenotype. Tracking the CD49e(low) expression under suspension culture conditions provides a mechanism to isolate an expanding suspension-grown MSC population with osteogenic potential. This could provide a potential strategy to isolate populations of MSCs, with functional osteogenic capacity, in a scalable and controllable culture system for therapeutic applications.
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Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Osteoblastos
/
Células de la Médula Ósea
/
Diferenciación Celular
/
Integrina alfa5
/
Células Madre Mesenquimatosas
Límite:
Humans
Idioma:
En
Revista:
Biotechnol Bioeng
Año:
2007
Tipo del documento:
Article
País de afiliación:
Canadá
Pais de publicación:
Estados Unidos