# Cellulose Plant-Derived Scaffolds as a Tool for Myometrium Modeling

## TL;DR

This study introduces plant-derived cellulose scaffolds for culturing mouse myometrial cells, offering a new 3D model to study uterine diseases like fibroids and adenomyosis.

## Contribution

A novel 3D in vitro model using plant-derived cellulose scaffolds to mimic the structure and function of the myometrium.

## Key findings

- Green onion leaf scaffolds enabled unidirectional cell orientation similar to native myometrial tissue.
- Cells cultured on the scaffolds remained viable for 2.5 months and expressed key smooth muscle markers.
- A multilayered scaffold system maintained cell viability and marker expression, mimicking native tissue structure.

## Abstract

The myometrium is the smooth muscle layer of the uterus, whose dysfunctions are involved in various pathologies leading to infertility, such as adenomyosis and uterine fibroids. Developing relevant in vitro models of the myometrium is crucial for investigating the pathogenesis of these diseases. In this study, we propose a novel approach for cultivating mouse 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, mimicking the tissue-specific organization of mouse myometrial SMCs in vivo. The mouse myometrial 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, connexin-43). To reproduce the native myometrium structure, a multilayered cultivation system was created. 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 modeling to study the pathogenesis of myometrium-associated diseases.