Cellulose Plant-Derived Scaffolds as a Tool for Myometrium Modelling

The myometrium is the smooth muscle layer of the uterus, whose dysfunctions are in-volved in various pathologies leading to infertility, such as adenomyosis and uterine fi-broids. Developing relevant in vitro models of the myometrium is crucial for investigat-ing the pathogenesis of these diseases. In this study, we propose a novel approach for cul-ti-vating myometrial smooth muscle cells (SMCs) using plant-derived cellulose scaffolds. The scaffolds were obtained through the decellularization of green onion leaf, celery stalk, or bluegrass leaf, subsequently coated with collagen type I. We found that the structure of the green onion leaf scaffold provides unidirectional orientation of cultured cells, mim-icking the tissue-specific organization of myometrial SMCs in vivo. The mouse myometri-al SMCs, cultured on this scaffold, proliferated, maintained viability up to 2.5 months, and retained the expression of the main markers of smooth muscle contractility (α-smooth muscle actin, transgelin, calponin, smooth muscle myosin heavy chains, con-nexin-43). To reproduce the native myometrium structure, a multilayered cultivation sys-tem was creat-ed. In a system of two overlaying scaffolds, cells also retained the viability and expression of smooth muscle contractility markers. The developed approach can be used for three-dimensional myometrium modelling to study the pathogenesis of myome-trium-associated diseases.