Enhancing Maturation of Human Neuromuscular Organoids via Electrical Stimulation

Area of Science:

• Stem cell biology
• Developmental biology
• Neuroscience

Background:

• Human pluripotent stem cell-derived organoids are valuable research models.
• Limited cellular maturity in organoids restricts their utility for studying adult physiology and diseases.

Purpose of the Study:

• To develop a method for enhancing the maturation of human neuromuscular organoids (NMOs).
• To investigate the effects of chronic Electrical Pulse Stimulation (EPS) on NMO development and function.

Main Methods:

• Applied low-frequency Electrical Pulse Stimulation (EPS) to developing NMOs over several weeks.
• Utilized quantitative imaging, transcriptomic analysis, and mechanobiological measurements to assess maturation.
• Compared EPS-trained NMOs (EPS-NMOs) with unstimulated controls.

Main Results:

• EPS significantly enhanced neuromuscular maturation and functional output in NMOs.
• EPS-NMOs exhibited stronger, more frequent spontaneous contractions and improved tissue morphology.
• Transcriptomic data revealed coordinated regulation of maturation-associated gene programs.
• Mechanobiological tests showed increased tissue stiffness and faster relaxation dynamics in EPS-NMOs.

Conclusions:

• Chronic EPS is an effective non-invasive strategy for promoting the morphological and functional maturation of human neuromuscular organoids.
• EPS-NMOs represent a more physiologically relevant model for studying neuromuscular development and disease.
• This approach offers an on-demand method to advance organoid complexity for research applications.