Abstract
Head and neck squamous cell carcinoma is a highly malignant disease and research is needed to find new therapeutic approaches. Faithful experimental models are required for this purpose. Here, we describe the specific cell culture conditions enabling the efficient establishment of primary cell culture models. Whereas a classical 10% serum-containing medium resulted in the growth of fibroblast-like cells that outcompeted epithelial cells, we found that the use of specific culture conditions enabled the growth of epithelial tumor cells from HPV+ and HPV− head and neck cancer tissue applicable for research. EpCAM and high Thy-1 positivity on the cell surface were mutually exclusive and distinguished epithelial and fibroblast-like subpopulations in all primary cultures examined and thus can be used to monitor stromal contamination and epithelial cell content. Interestingly, cells of an individual patient developed tumor spheroids in suspension without the use of ultra-low attachment plates, whereas all other samples exclusively formed adherent cell layers. Spheroid cells were highly positive for ALDH1A1 and hence displayed a phenotype reminiscent of tumor stem cells. Altogether, we present a system to establish valuable primary cell culture models from head and neck cancer tissue at high efficiency that might be applicable in other tumor entities as well.
Highlights
Head and neck cancer is a highly malignant disease with 600,000 new incidences worldwide per year and a mortality rate of about 50% [1,2]
Primary cell cultures derived from surgically excised human tumor tissue are highly valuable research tools to investigate the biology and therapeutic vulnerability of cancer cells considering the genetics of each individual neoplasm
This shows that classical DMEM + 10% fetal bovine serum (FBS) is neither suitable to expand head and neck squamous cell carcinomas (HNSCCs) cells with epithelial morphology in vitro nor to preserve phenotypically undifferentiated cell populations within cultures
Summary
Head and neck cancer is a highly malignant disease with 600,000 new incidences worldwide per year and a mortality rate of about 50% [1,2]. About 90% of all tumors in the head and neck area are head and neck squamous cell carcinomas (HNSCCs) and occur in heterogeneous locations. They arise from the mucosal epithelium of the oral cavity, larynx, and pharynx [3]. HNSCCs in oropharyngeal location can be subdivided into cases caused by the human papilloma virus (HPV) and those cases that are HPV-negative, which differ strikingly in their molecular biology [4]. HPV-negative oropharyngeal HNSCCs have a worse prognosis than HPV-positive tumors [7], and the survival of HPV-negative HNSCC patients has not improved substantially in recent decades [8], so new therapeutic approaches are needed to treat this malignancy
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