Orosphere assay: A method for propagation of head and neck cancer stem cells
Article first published online: 13 JUL 2012
Copyright © 2012 Wiley Periodicals, Inc.
Head & Neck
Volume 35, Issue 7, pages 1015–1021, July 2013
How to Cite
Krishnamurthy, S. and Nör, J. E. (2013), Orosphere assay: A method for propagation of head and neck cancer stem cells. Head Neck, 35: 1015–1021. doi: 10.1002/hed.23076
- Issue published online: 20 JUN 2013
- Article first published online: 13 JUL 2012
- Manuscript Accepted: 23 APR 2012
- Weathermax Foundation, University of Michigan Comprehensive Cancer Center
- National Institutes of Health. Grant Numbers: P50-CA-97248 (University of Michigan Head and Neck SPORE), R01-DE15948, R21-DE19279, R01-DE21139
- squamous cell carcinoma;
- suspension culture;
Recent evidence suggests that head and neck squamous cell carcinomas (HNSCCs) harbor a small subpopulation of highly tumorigenic cells, designated cancer stem cells. A limiting factor in cancer stem cell research is the intrinsic difficulty of expanding cells in an undifferentiated state in vitro.
Here, we describe the development of the orosphere assay, a method for the study of putative head and neck cancer stem cells. An orosphere is defined as a nonadherent colony of cells sorted from primary HNSCCs or from HNSCC cell lines and cultured in 3-dimensional soft agar or ultralow attachment plates. Aldehyde dehydrogenase activity and CD44 expression were used here as stem cell markers.
This assay allowed for the propagation of head and neck cancer cells that retained stemness and self-renewal.
The orosphere assay is well suited for studies designed to understand the pathobiology of head and neck cancer stem cells. © 2012 Wiley Periodicals, Inc. Head Neck, 2013