INTRODUCTION: The small number of cancer stem cells, which correspond to only 0.01% - 0.1% of total tumor cells, has been the biggest obstacle in understanding their biology and role in the origin and maintenance of tumors, their metastatic and recurrence potentials, and resistance to radio-chemotherapy. Therefore, promoting its accumulation will enable further studies and future advances in the diagnosis and treatment of head and neck cancer squamous cell carcinoma. OBJECTIVE: To induce cancer stem cell accumulation in primary cell cultures of head and neck squamous cell carcinoma using a hypoxia chamber. METHODS: Head and neck squamous cell carcinoma samples were cultured and subjected to hypoxia. Oxygen deprivation aimed to induce cancer stem cell accumulation. RESULTS: Immediately after hypoxia, the percentage of O2-deprived cancer stem cells increased 2-fold as compared to control. Surprisingly, new phenotyping performed 45 days after hypoxia showed a 9-fold increase in cancer stem cell percentage in cells that suffered hypoxia. Hypoxic cells showed an increase in spheroid formation when compared to control cells, as well as enhanced abilities in invasion and migration. CONCLUSION: Hypoxia was efficient in cancer stem cell accumulation. As cancer stem cells are a small number of cells within the tumor, promoting their accumulation will enable further studies and future advances in the diagnosis and treatment of head and neck cancer.
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