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Orosphere assay: a method for propagation of head and neck cancer stem cells.


ABSTRACT: BACKGROUND: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. METHODS: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. RESULTS:This assay allowed for the propagation of head and neck cancer cells that retained stemness and self-renewal. CONCLUSION:The orosphere assay is well suited for studies designed to understand the pathobiology of head and neck cancer stem cells.

SUBMITTER: Krishnamurthy S 

PROVIDER: S-EPMC3887391 | biostudies-literature | 2013 Jul

REPOSITORIES: biostudies-literature

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Orosphere assay: a method for propagation of head and neck cancer stem cells.

Krishnamurthy Sudha S   Nör Jacques E JE  

Head & neck 20120713 7


<h4>Background</h4>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.<h4>Methods</h4>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  ...[more]

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