Three-Dimensional Organoids Culturing and Processing Techniques for Myometrial and Uterine Fibroids Generation

Uterine fibroids are the most common benign, monoclonal, gynecological tumors in a woman's uterus. To overcome the common obstacles related to the methods used in studying these pathologies, we aimed to devise a strategy to generate three-dimensional organoid models of myometrium and uterine fibroid stem cells using collagen-containing hydrogels as embedding scaffolds. Specifically, collagen-containing hydrogels in a low attachment V-bottom 96-well plates were exploited. This method allowed the development of 3D organoids of two stem cell types from normal myometrium and uterine fibroid-containing myometrium by embedding them in collagen-containing hydrogels and forming organoids in a suitable stem cell proliferation medium. The organoids successfully differentiated, proliferated, and self-organized into complex structures, developing a sustainable system. The importance of this model is the understanding of pathophysiology and etiopathogenesis, as well as for testing new drugs to prevent or treat uterine fibroids.