Formation and Long-Term Culture of hiPSC-Derived Sensory Nerve Organoids Using Microfluidic Devices

Area of Science:

• Stem Cell Biology
• Neuroscience
• Bioengineering

Background:

• Established methods exist for motor nerve organoids derived from human induced pluripotent stem cells (hiPSCs).
• Generating sensory nerve organoids from hiPSCs remains a challenge.
• Sensory neurons are crucial for detecting stimuli like pain and temperature.

Purpose of the Study:

• To investigate the feasibility of generating sensory nerve organoids using hiPSC-derived sensory neurons.
• To develop a microfluidic approach for unidirectional axon elongation and long-term culture.
• To assess the functional properties of the generated sensory nerve organoids.

Main Methods:

• Utilized I-shaped microchannels in polydimethylsiloxane (PDMS) chips for unidirectional axon growth.
• Cultured hiPSC-derived neurospheres to form sensory nerve organoids with axon bundles up to 6 mm long within 14 days.
• Maintained organoid cultures for over 60 days through regular medium exchange.

Main Results:

• Successfully generated long axon bundles (over 6 mm) from hiPSC-derived sensory neurons within 14 days.
• Achieved long-term culture viability for the sensory nerve organoids exceeding 60 days.
• Confirmed the presence of sensory neuron markers (ISL1) and functional TRPV1 receptors, evidenced by calcium influx upon capsaicin stimulation.

Conclusions:

• Demonstrated the feasibility of creating functional sensory nerve organoids using a microfluidic system.
• Highlighted the potential of these organoids for long-term culture and applications in bioengineered nociceptive sensors.
• Paved the way for future research in sensory nervous system modeling and drug screening.