MSLASpheroidStamp: 3d cell spheroids for everyone

3D cell cultures, such as cell spheroids, are actively used in biology for modeling biological processes, studying intercellular interactions, and screening pharmacological compounds and are becoming indispensable objects in cell culture laboratories. There are many methods for producing spheroids, which vary in cost and convenience. One of the most convenient and affordable methods is the use of agarose microwells. We developed approaches to fabricate agarose microwells in standard culture plastic with the assistance of a hobby-grade MSLA 3D printer. The use of 3D printing allows the customization of microwells in a wide range of shapes and sizes and scales the production process from a few spheroids to tens of thousands. We have shown that it is possible to create gel microwells in a dish with a glass bottom, which allows us to easily realize time-lapse confocal microscopy of spheroids as well as in situ optical clearing in the same dishes to study the spheroid structure. We demonstrated the ability to study the cytotoxicity of various substances and nanoparticles in commonly used 96-well plates.

Finally, in this article, we describe the difficulties and limitations of our approach and suggest ways to solve them, allowing the reader not only to reproduce it, but also to adapt it to the specific needs of a certain laboratory, using the provided 3D models and instructions.