Document Type : Protocol
Authors
1
Department of Human Genetics, Faculty of Biomedical Sciences and Technology, Sri Ramachandra Institute of Higher Education and Research, Porur, Chennai, Tamil Nadu, India.
2
Department of Obstetrics and Gynaecology, Saveetha Medical College and Hospital, Saveetha Institute of Medical and Technical Sciences, Saveetha Nagar, Thandalam, Chennai, Tamil Nadu, India.
3
Department of Research, Madha Medical College and Research Institute, Kundrathur Main Road, Kovur, Thandalam, Chennai, Tamil Nadu, India.
Abstract
Objective: Human cancer cell lines have contributed immensely to cancer research. Advancements in cell culture techniques and contributions from modern biomedical engineering developments have enabled better utilization of such cell lines. There has been a major shift from the two-dimensional (2D) way of cell culture to the three-dimensional (3D) ways of culturing cells, which began about two decades ago. We propose an extension to this evolving trend in the form of 3D reverts (3DRs). Methods: 3D aggregates of three human cancer cell lines, HCT-116, MG-63, and SiHa, were obtained using agarose hydrogels as the matrix. 3DRs were obtained by introducing the floating 3D aggregates into scaffold-free culture units. These 3DRs were observed periodically, and images were obtained and analyzed for their culture characteristics. Results: The 3DRs of the three cell lines mimicked an explant-like features, with cells migrating out of the aggregates and attaching to the culture surfaces. Each cell line exhibited a unique pattern of migration of individual cells from their respective 3D aggregates to form 3DRs. The cells in the proliferative zone of HCT-116, MG-63, and SiHa aggregates showed single-cell mesenchymal-like, amoeboid-like and collective migration, respectively. The morphological features of the 3D aggregates of the cell lines used largely determined the type of cell movement exhibited for formation of the 3DRs. Conclusions: 3DR types of culture have not been well studied or described in detail previously. Although such cultures resemble 2D monolayers, the manner in which they develop differs among cell lines. Such 3DRs have potential as emerging models for in vitro cancer research experiments, especially for studies related to metastasis.
Keywords
Main Subjects
)