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Endothelial cell behavior inside myoblast sheets with different thickness
Trung Xuan Ngo1, Eiji Nagamori, Tetsutaro Kikuchi
1Division of Chemical Engineering, Graduate School of Engineering Science, Osaka University, Toyonaka, Osaka, Japan.
The thickness of human skeletal muscle myoblast (HSMM) sheets influences endothelial cell behavior. Varying HSMM layer numbers alters how human umbilical vein endothelial cells (HUVECs) migrate and aggregate within the construct.
Area of Science:
- Tissue Engineering
- Cell Biology
- Biomaterials Science
Background:
- Endothelial cell behavior is crucial for vascularization in tissue engineering.
- Understanding cell-cell interactions and migration within engineered tissues is vital for functional construct development.
Purpose of the Study:
- To investigate the impact of human skeletal muscle myoblast (HSMM) sheet thickness on the behavior of green fluorescent protein expressing human umbilical vein endothelial cells (GFP-HUVECs).
- To establish an in vitro culture system for observing endothelial cell dynamics within a layered myoblast construct.
Main Methods:
- Developed an in vitro cell sheet stacking system using GFP-HUVECs and HSMM sheets.
- Cultured GFP-HUVECs under HSMM sheets with varying layer numbers (monolayer, 3, 5, and 7 layers).
- Incubated cultures for 96 hours and observed HUVEC migration, localization, and aggregate morphology.
Main Results:
- Monolayer HSMM: HUVECs rapidly migrated to the top and detached.
- 3-layered HSMM: HUVECs migrated to the top layer, forming island aggregates.
- 5-layered HSMM: HUVECs migrated to the middle, forming net-shaped aggregates.
- 7-layered HSMM: HUVECs migrated to the basal layer, forming sparse net-shaped aggregates.
Conclusions:
- HSMM sheet thickness, controlled by layer number, is a critical parameter affecting HUVEC migration time, encounters, localization, and morphology.
- This study demonstrates the influence of the microenvironment's physical properties on endothelial cell behavior in a 3D engineered tissue context.
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