Elucidation and regulation of heterogeneous cell behavior inside a multilayered skeletal muscle myoblast sheet

Abstract

Tissue organization is governed by self-assembly phenomenon. In tissue architecture, variety of heterogeneous cell population habitat autonomously in the proper regions and maintain their organization. In our previous studies, a multi-layered human skeletal muscle myoblast (HSMM) sheet regarded as a plate-shaped aggregate was prepared and co-incubated on heterogeneous cell population, such as human umbilical vein endothelial cells (HUVECs), human skeletal muscle fibroblasts (HSMFs) or human mesenchymal stem cells (HMSCs) etc., to quantitatively elucidate their behavior inside the HSMM tissue. Stereoscopic observation by confocal laser scanning microscopy with image processing revealed random or collective migration and habitat aggregation of heterogeneous cell population inside the HSMM sheet depending on their character of cell-cell connection and migration. After understanding heterogeneous cell behavior inside the HSMM sheet, modified method to construct multilayered HSMM sheet with uniform HUVEC network by stacking single HSMM sheets including pre-formed 2D HUVEC network was successfully established.