Abstract
Cancer stem cells from oropharyngeal squamous cell carcinoma (OPSCC) have the ability to self-renew and differentiate into heterogeneous three-dimensional structures carrying features of tumor cells. Here, we describe a simple and label-free method for generating tumor organoids, and imaging them using live digital holographic microscopy (DHM) on the basis of the phase shift caused by light passing through the cells. We show early events of cell aggregation during tumor-organoid formation, and display their heterogeneity in terms of optical parameters up to an optical volume of 105 µm3. Lastly, by sorting OPSCC epithelial cells, we demonstrate that CD44+ cells displayed greater motility and tumor-forming capacity than those of CD44− cells. These results were in line with previous reports highlighting increased invasive and tumorigenic potential in tumor cells expressing high levels of CD44. Our method provides insight into the formation of tumor organoids, and could be used to assess stemness-associated biomarkers and drug screenings on the basis of tumor organoids.
Highlights
Oropharyngeal squamous cell carcinoma (OPSCC) is a head-and-neck cancer affecting the tonsils, base of the tongue, pharyngeal walls, and soft-palate region [1]
When cultured in a hydrogel, cancer stem cells (CSCs) generate 3D structures known as organoids, self-organized cellular structures that exhibit phenotypes resembling those seen in tumors [3,4]
48-year-old nonsmoking man diagnosed with poorly differentiated tonsillar squamous-cell carcinoma (Stage T2N0M0) was previously described [13]
Summary
Oropharyngeal squamous cell carcinoma (OPSCC) is a head-and-neck cancer affecting the tonsils, base of the tongue, pharyngeal walls, and soft-palate region [1]. When cultured in a hydrogel, CSCs generate 3D structures known as organoids, self-organized cellular structures that exhibit phenotypes resembling those seen in tumors [3,4]. Basement-membrane matrices provide appropriate mechanical stimuli that allow CSCs embedded within them to aggregate, grow, and differentiate, resulting in organoids of different sizes and morphologies [4,5] with the ability to replicate cellular, mechanical, and physical signals that take place in the tumor microenvironment [6,7]. In OPSCC, CD44 and nerve growth-factor receptor (NGFR) are two candidate markers for identifying cancer stem cells [8,9], as shown by a study where CD44+ /NGFR+
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