A043 - A practical review of several 3D-culture methods for the generation of hollow or solid organoids/spheroids with a unique cell-type, how environment matters

Laetitia Andrique (laetitia.andrique@u-bordeaux.fr)
Alessandro Furlan (alessandro.furlan@univ-lille.fr)

In this workshop we will illustrate how using several methods for generating organoids influences the final topology, histology and differentiation of cells. As a substitute for hiPSCs that, for obvious safety reasons we cannot use in the context of MiFoBio, we will take advantage of a versatile alternative, i.e., an immortalised epithelial human cell line. The attendees will implement different methods used to generate 3D aggregates of cells, in the cell culture room. The different methods differ in many ways, both in the generation process and the final cellular topology. We will follow the growth of cell aggregates, in time and provide to the attendees the images of the structures that will illustrate how they are different in terms of initial state and growth. The participants will as well carry out some methods to label and monitor cell growth by using fluorescent dyes. Finally, we will show how to prepare and mount the different samples for microscopy depending on their topology, providing tips and tricks that would help the attendees to set-up their own experiments back in their labs.