Three-Dimensional Cell Culture Models to Investigate Oral Carcinogenesis: A Scoping Review

Abstract

Three-dimensional (3-D) cell culture models, such as spheroids, organoids, and organotypic cultures, are more physiologically representative of the human tumor microenvironment (TME) than traditional two-dimensional (2-D) cell culture models. They have been used as in vitro models to investigate various aspects of oral cancer but, to date, have not be widely used in investigations of the process of oral carcinogenesis. The aim of this scoping review was to evaluate the use of 3-D cell cultures in oral squamous cell carcinoma (OSCC) research, with a particular emphasis on oral carcinogenesis studies. Databases (PubMed, Scopus, and Web of Science) were systematically searched to identify research applying 3-D cell culture techniques to cells from normal, dysplastic, and malignant oral mucosae. A total of 119 studies were included for qualitative analysis including 53 studies utilizing spheroids, 62 utilizing organotypic cultures, and 4 using organoids. We found that 3-D oral carcinogenesis studies had been limited to just two organotypic culture models and that to date, spheroids and organoids had not been utilized for this purpose. Spheroid culture was most frequently used as a tumorosphere forming assay and the organoids cultured from human OSCCs most often used in drug sensitivity testing. These results indicate that there are significant opportunities to utilize 3-D cell culture to explore the development of oral cancer, particularly as the physiological relevance of these models continues to improve.

New Scenarios in Pharmacological Treatments of Head and Neck Squamous Cell Carcinomas

“…Traditional drug testing performed in 2D cultures allowed us to have a better understanding of the potential effects and toxicity of the new drugs. However, 2D cultures only partially replicate the complex environment found in patients, and, thus, the use of more appropriate culture model systems is essential for drug testing [ 68 , 69 , 70 ].…”

“…Three-dimensional (3D) culture models (e.g., organotypic cultures, spheroids, and organoids) are more similar to the native tumor regarding cell heterogeneity, genetic variability, and cell to microenvironment interaction, thus representing an excellent platform for personalized medicine and drug screening [ 68 , 71 ].…”

“…In general, spheroids contain a proliferating cell layer (external layer) and a quiescent cell layer (internal layer) [ 71 , 72 , 73 , 74 , 75 , 76 , 77 , 78 , 79 , 80 , 81 , 82 , 83 , 84 , 85 , 86 ]. Various techniques can be used to grow and maintain spheroids in culture, with applications ranging from studies on tumor microenvironment to drug screening and molecular testing [ 68 , 78 , 87 , 88 ]. In one of the most common methods for spheroid production, the hanging drop, a drop of culture medium containing patient’s derived cells, is plated, suspended, and cells aggregate by gravity at the bottom of the drop.…”

“…Organoids can also be used to study TME and drug screening [ 101 , 102 ]. They can be cultured in a matrix, or in suspension, with specific supplemented media [ 68 , 101 , 103 ]. Organoids can be expanded from patient-derived cells and maintained in culture for long periods, allowing the development of personalized biobanks [ 68 ].…”

“…They can be cultured in a matrix, or in suspension, with specific supplemented media [ 68 , 101 , 103 ]. Organoids can be expanded from patient-derived cells and maintained in culture for long periods, allowing the development of personalized biobanks [ 68 ]. Cancer-derived organoids are able to preserve the characteristics of the original tumor [ 101 , 104 , 105 , 106 ], including tumor metabolism [ 68 , 107 ], and, therefore, constitute ideal tools for identifying novel cancer-specific biomarkers [ 108 , 109 ].…”