Generating human skeletal myoblast spheroids for vascular myogenic tissue engineering

Area of Science:

• Biomedical Engineering
• Tissue Engineering
• Regenerative Medicine

Background:

• Engineered myogenic microtissues from human skeletal myoblasts are valuable for in vitro applications.
• Incorporating vascular structures into these models presents a significant challenge.

Purpose of the Study:

• To develop a high-throughput method for generating complex, vascularized skeletal muscle microtissues.
• To create advanced 3D models for in vitro drug testing and disease modeling.

Main Methods:

• Generated myogenic spheroids using high-throughput, non-adhesive micropatterned surfaces.
• Co-cultured human skeletal myoblasts with human adipose-derived stem cells to improve spheroid integrity and myogenic properties.
• Incorporated endothelial cells to create pre-vascularized microtissues with capillary-like networks.
• Demonstrated spheroid assembly into larger constructs via doublet fusion.

Main Results:

• Achieved uniform, viable co-culture spheroids with enhanced myogenic properties at an optimal cell ratio.
• Successfully formed vascularized spheroids containing capillary-like networks, lumina, and extracellular matrix components.
• Confirmed retention of myogenicity and endothelial cell sprouting capabilities.
• Proved the ability of spheroids at different maturation stages to assemble into larger constructs.

Conclusions:

• Developed a robust, high-throughput method for generating complex, vascularized skeletal muscle microtissues.
• The engineered microtissues possess tissue-specific microarchitecture suitable for advanced in vitro applications.
• These 3D microtissues are a promising tool for in vitro drug testing and human disease modeling.