MSLASpheroidStamp: 3d cell spheroids for everyone

Abstract 3D cell cultures, such as cell spheroids, are actively used in biology for modeling biological processes, studying cell-cell interactions and screening various drugs and are becoming indispensable objects in cell culture laboratories. There are many methods for producing spheroids, varying in cost and convenience. One of the most handy and affordable is the use of agarose microwells. We have 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 you to customize microwells in a wide range of shapes and sizes, and scale the production process from a few spheroids to tens of thousands. We demonstrated the ability to cultivate spheroids in a glass bottom dish and dynamically observe their formation, perform in situ optical clearing, and demonstrated the ability to study the cytotoxicity of various substances and nanoparticles in 96-well plates. And finally, in this article we describe the difficulties and limitations of our approach and suggest ways for solving them, allowing you not only to reproduce it, but also to adapt it to the specific needs of your laboratory, using 3D models and our instructions.