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Fully Human Tumor-based Matrix in Three-dimensional Spheroid Invasion Assay
Published on: May 7, 2019
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Fully Human Tumor-based Matrix in Three-dimensional Spheroid Invasion Assay
Erika Naakka1, Katja Tuomainen1, Henrik Wistrand1
1Department of Oral and Maxillofacial Diseases, Clinicum, Faculty of Medicine, University of Helsinki.
Journal of Visualized Experiments : Jove
|May 28, 2019
Summary
This study introduces a 3D human tumor-based spheroid invasion assay using a human myoma-derived matrix (HMDM). This novel method offers a more reliable in vitro model for studying cancer cell invasion compared to traditional assays.
Area of Science:
- Oncology
- Biotechnology
- Cell Biology
Background:
- Two-dimensional (2D) cell cultures lack crucial tumor microenvironment elements, limiting in vitro cancer research reliability.
- Three-dimensional (3D) cell culture assays improve reliability by enabling cancer cell interaction with the extracellular matrix.
- Existing 3D spheroid invasion assays primarily use animal-derived matrices, which may not accurately mimic the human tumor microenvironment.
Purpose of the Study:
- To develop and validate a simple, reproducible, and reliable 3D spheroid invasion assay using a human-derived matrix.
- To establish a 3D in vitro model that better simulates the human tumor microenvironment for studying cancer cell invasion.
- To provide detailed protocols for imaging and analysis of cancer cell invasion in a 3D human matrix.
Main Methods:
- Development of a human myoma-derived matrix (HMDM)/fibrin matrix for 3D cell culture.
- Culturing cancer cell spheroids in ultra-low attachment plates within the HMDM/fibrin matrix.
- Daily imaging and analysis of cancer cell invasion using ilastik and Fiji ImageJ software.
Main Results:
- The HMDM/fibrin matrix demonstrated superior induction of carcinoma cell migration and invasion compared to animal-derived matrices (e.g., MSDM).
- The developed assay provides a robust platform for studying cancer cell invasion in a more physiologically relevant 3D human context.
- The protocol was successfully demonstrated using human laryngeal squamous cell carcinoma cell lines, indicating broad applicability to other solid cancers.
Conclusions:
- The HMDM/fibrin matrix-based 3D spheroid invasion assay is a valuable tool for in vitro cancer research.
- This human-based 3D model enhances the study of cancer cell invasion, offering more reliable insights than traditional methods.
- The protocol's simplicity, reproducibility, and suitability for various solid cancers make it a significant advancement in cancer research methodology.
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