Method for Culturing and Imaging Homogeneous 3D Cancer Spheroids as a Model for the Uptake of DNA Nanostructures

Owing to their natural biocompatibility, structural stability, ease of internalization, and editable functionality, DNA nanostructures show promising potential as drug candidates and potential vehicles for drug delivery. 3D cell culture is currently recognized as a very useful technique to recapitulate the behavior of cancer and its microenvironment and test the results of drug treatment with higher predictable value than the most common and traditional 2D cell models. Techniques are emerging to make the culturing of 3D models easy and statistically robust as a tool aiding drug development. In this chapter, we present an efficient and reproducible method to generate homogeneous cancer spheroids that can be used to characterize the uptake of nanostructured drugs, such as DNA nanostructure. Moreover, we describe the procedure to obtain a structural characterization of such processes by lightsheet fluorescence microscopy technology.