A non-contact suspension culture approach to the culture of osteogenic cells derived from a CD49elow subpopulation of human bone marrow-derived cells
✍ Scribed by Dolores Baksh; Peter W. Zandstra; John E. Davies
- Publisher
- John Wiley and Sons
- Year
- 2007
- Tongue
- English
- Weight
- 490 KB
- Volume
- 98
- Category
- Article
- ISSN
- 0006-3592
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✦ Synopsis
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 CFUosteoblast (CFU-O) activity was associated with a CD45 À CD49e low phenotype. Importantly, suspension-grown MSCs, capable of CFU-F and CFU-O development, maintained the CD45 À CD49e low phenotype whereas MSCs directly cultured under adherent conditions rapidly upregulated CD49e expression and were associated with a CD45 À 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.