The video describes the laser dissection of the 3D organotypic co-culture model with micron-level precision. In 3D organotypic models, stromal constituents such as fibroblasts are 3-dimensionally co-cultured with cancer epithelial cells. The 3D model serves as an experimental tool that enables invasion and cancer-stroma interactions to be studied in near-physiological conditions.
Protocol
1. Organotypic Preparation: Preparation of the fibroblast impregnated organotypic gel: Prepare a fibroblast cell suspension containing 5 x 105 cells per organotypic in DMEM supplemented with 10% FCS and 292 μg/ml L-Glutamine. Make up the organotypic gels on ice in the following ratios: • 7 volumes of Rat-Tail Collagen: Matrigel mix (1:1) • 1 volume of filtered 10x DMEM …
Declarações
The authors have nothing to disclose.
Materials
Colorectal cancer cell lines – example shown SW480
Laser Microdissection: A Technique to Isolate Invading Colorectal Cancer cells from a 3D Organotypic Culture Model. J. Vis. Exp. (Pending Publication), e20343, doi: (2023).