Generation of Organoids and Analysis of Ferroptosis in Organoids

Ferroptosis is a unique form of iron-dependent cell death induced by lipid peroxidation and subsequent plasma membrane rupture, which sets it apart from other types of regulated cell death. Ferroptosis has been linked to a diverse range of biological processes, such as aging, immunity, and cancer. Organoids, on the other hand, are three-dimensional (3D) miniaturized model systems of different organs in vitro cultures, which have gained widespread interest for modeling tissue development and disease, drug screening, and cell therapy. Organoids offer tremendous potential for improving our understanding of human diseases, particularly in the search for the field of ferroptosis in pathological processes of organs. Furthermore, cancer organoids are utilized to investigate molecular mechanisms and drug screening in vitro due to the anti-tumor effect of ferroptosis. Currently, the development of liver organoids has reached a relatively mature stage. Here, we present the protocols for the generation of liver organoids and liver cancer organoids, along with the methods for detecting ferroptosis in organoids.