Human Decellularized Endometrial Scaffold Seeded by Myometrial Smooth Muscle Cells Supports the Differentiation of Human Endometrial Mesenchymal Cells to Epithelial and Stromal Cells
Human Decellularized Endometrial Scaffold Seeded by Myometrial Smooth Muscle Cells Supports the Differentiation of Human Endometrial Mesenchymal Cells to Epithelial and Stromal Cells
Introduction: This study designed to evaluate the co-culturing of human mesenchymal endometrial cells (EMCs) and myometrial smooth muscle cells (MSMCs) in decellularized scaffold as a natural bioscaffold to formation of the endometrial-like structure.
Methods: Following decellularization of the human endometrium, Human mesenchymal endometrial cells were seeded onto decellularized human endometrium using centrifugation at different speeds and times, resulting in 15 experimental subgroups. We analyzed the number of residual cell count in suspension in all subgroups and selected the method with the lower number of suspended cells for further investigation. Then, the human endometrial mesenchymal cells and the myometrial muscle cells were seeded on the decellularized tissue and cultured for 1 wk; then, differentiation of the seeded cells was assessed by morphological and gene expression analysis.
Results: Centrifugation at 6020 g for 2 minutes proved to be the most effective cell seeding method, resulting in the number of seeded cells and the lowest number of residual cells in suspension. The recellularized scaffold displayed endometrial-like cells with surface protrusions, while the stromal cells exhibited spindle and polyhedral morphology. The myometrial cells almost were homed at the periphery of the scaffold and mesenchymal cells penetrated in deeper parts similar to their arrangement in the native uterus.The more expression of endometrial-related genes such as SPP1, MMP2, ZO-1, LAMA2, and COL4A1 and low-level expression of the OCT4 gene as a pluripotency marker confirmed the differentiation of seeded cells.
Conclusion: Endometrial-like structures were formed by the co-culturing of human endometrial mesenchymal cells and smooth muscle cells on the decellularized endometrium