4.7 Article

Microstructured soft devices for the growth and analysis of populations of homogenous multicellular tumor spheroids

Journal

CELLULAR AND MOLECULAR LIFE SCIENCES
Volume 80, Issue 4, Pages -

Publisher

SPRINGER BASEL AG
DOI: 10.1007/s00018-023-04748-1

Keywords

Microwells; Microtissues; Spheroid; Cancer models; 3D cell culture; Drug screening; High content screening; High-throughput screening

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Multicellular tumor spheroids are an improved in vitro model used for cancer biology research and drug screening. A simple method has been developed to create custom microwell arrays made of polydimethylsiloxane or agarose, providing a tunable mechanical environment. Various cancer and non-cancer cell lines can be cultured in these devices, and the growth, viability, and response to treatments can be characterized using molecular and imaging techniques. This platform enables statistically robust investigations and confocal microscopy imaging while retaining the geographical arrangement of spheroids.
Multicellular tumor spheroids are rapidly emerging as an improved in vitro model with respect to more traditional 2D culturing. Microwell culturing is a simple and accessible method for generating a large number of uniformly sized spheroids, but commercially available systems often do not enable researchers to perform complete culturing and analysis pipelines and the mechanical properties of their culture environment are not commonly matching those of the target tissue. We herein report a simple method to obtain custom-designed self-built microwell arrays made of polydimethylsiloxane or agarose for uniform 3D cell structure generation. Such materials can provide an environment of tunable mechanical flexibility. We developed protocols to culture a variety of cancer and non-cancer cell lines in such devices and to perform molecular and imaging characterizations of the spheroid growth, viability, and response to pharmacological treatments. Hundreds of tumor spheroids grow (in scaffolded or scaffold-free conditions) at homogeneous rates and can be harvested at will. Microscopy imaging can be performed in situ during or at the end of the culture. Fluorescence (confocal) microscopy can be performed after in situ staining while retaining the geographic arrangement of spheroids in the plate wells. This platform can enable statistically robust investigations on cancer biology and screening of drug treatments.

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