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Updated: May 7, 2026
Longitudinal Morphological and Physiological Monitoring of Three-dimensional Tumor Spheroids Using Optical Coherence Tomography
Published on: February 9, 2019
Characterization of squamous cell carcinoma in an organotypic culture via subsurface non-linear optical molecular
Christina S Scanlon1, Elizabeth A Van Tubergen, Leng-Chun Chen
1Department of Periodontics and Oral Medicine, School of Dentistry, University of Michigan, MI 48109-1078, USA.
Abstract:
Tristetraprolin (TTP) is an RNA-binding protein which downregulates multiple cytokines that mediate progression of head and neck squamous cell carcinoma (HNSCC). We previously showed that HNSCC cells with shRNA-mediated knockdown of TTP are more invasive than controls. In this study, we use control and TTP-deficient cells to present a novel subsurface non-linear optical molecular imaging method using a three-dimensional (3D) organotypic construct, and compare the live cell imaging data to histology of fixed tissue specimens. This manuscript describes how to prepare and image the novel organotypic system that closely mimics HNSCC in a clinical setting. The oral cancer equivalent (OCE) system allows HNSCC cells to stratify and invade beyond the basement membrane into underlying connective tissue prepared from decellularized human dermal tissue. The OCE model was inspired by tissue engineering strategies to prepare autologous transplants from human keratinocytes. Advantages of this method over previously used in vitro cancer models include the simulation of the basement membrane and complex connective tissue in the construct, in addition to the ability to track the 3D movement of live invading cells and quantify matrix destruction over time. The OCE model and novel live cell imaging strategy may be applied to study other types of 3D tissue constructs.
Insights
Tristetraprolin (TTP) deficiency increases head and neck cancer invasion. A new 3D organotypic model and imaging method track live cancer cell invasion and matrix destruction, mimicking clinical HNSCC progression.
Area of Science:
- Biomedical Engineering
- Cancer Research
- Optical Imaging
Background:
- Tristetraprolin (TTP) downregulates cytokines driving head and neck squamous cell carcinoma (HNSCC) progression.
- TTP-deficient HNSCC cells exhibit increased invasiveness.
- Current in vitro models lack complex tissue microenvironments.
Purpose of the Study:
- To develop and validate a novel 3D organotypic construct for studying HNSCC invasion.
- To present a subsurface non-linear optical molecular imaging method for live cell tracking.
- To compare live imaging data with histology of fixed tissue specimens.
Main Methods:
- Preparation of a 3D oral cancer equivalent (OCE) model using decellularized human dermal tissue and keratinocytes.
- Utilizing subsurface non-linear optical molecular imaging to track live HNSCC cell invasion.
- Stratification and invasion of HNSCC cells beyond the engineered basement membrane into connective tissue.
Main Results:
- The OCE model successfully mimics HNSCC invasion beyond the basement membrane.
- Live cell imaging allowed real-time tracking of 3D cell movement and matrix degradation.
- The imaging data correlated with histology of fixed tissue specimens.
Conclusions:
- The OCE model provides a more clinically relevant in vitro system for HNSCC research.
- The novel imaging strategy enables quantification of matrix destruction by invading cancer cells.
- This approach can be adapted for studying other 3D tissue constructs and diseases.
