Protocol for formation, staining, and imaging of 3D breast cancer models using MicroTissues mold systems
Year: 2024
Type: article
Abstract: The use of 3D cancer models to study therapy response has a great advantage over conventional 2D cell culture. Here, we present an optimized protocol for breast cancer spheroid and rod-shaped microtissue formation using MicroTissues mold systems. We describe steps for cast formation and treating the 3D models with DNA-damaging agents. We then detail procedures for analyzing the 3D models by whole-mount immunostaining and confocal imaging of fixed samples or with the use of live-cell reporters. • Instructions for cast formation required to form spheroids and rods • Procedure to form spheroids and rods and to treat these models • Instructions for harvesting of 3D cell models and immunostaining of spheroids • Instructions for confocal imaging of live and fixed 3D cell models Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. The use of 3D cancer models to study therapy response has a great advantage over conventional 2D cell culture. Here, we present an optimized protocol for breast cancer spheroid and rod-shaped microtissue formation using MicroTissues mold systems. We describe steps for cast formation and treating the 3D models with DNA-damaging agents. We then detail procedures for analyzing the 3D models by whole-mount immunostaining and confocal imaging of fixed samples or with the use of live-cell reporters.
Source: STAR Protocols
Authors: Ben Haspels, Maayke M.P. Kuijten
Institutions: Erasmus MC, Erasmus MC Cancer Institute
Language: English
Subfield: Biomedical Engineering
Field: Engineering
Domain: Physical Sciences
Open Access status: gold
Estimated APC paid: $1,500
OpenAlex ID: W4402682523