MSLASpheroidStamp: 3d cell spheroids for everyone

Abstract

3D cell cultures, such as cell spheroids, are actively used in biology for modeling biological processes, studying intercellular interactions and pharmacological compounds screening 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 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, and we have also performed 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. And finally, in this article we describe the difficulties and limitations of our approach and suggest ways for solving them, allowing the reader not only to reproduce it, but also to adapt it to the specific needs of a certain laboratory, using provided 3D models and instructions.