Three-Dimensional Motor Nerve Organoid Generation

作者： Tatsuya Osaki1,2, Siu Yu A. Chow1,2, Yui Nakanishi1,2, Joel Hernández1,3, Jiro Kawada4, Teruo Fujii1, Yoshiho Ikeuchi1,2 1Institute of Industrial Science,The University of Tokyo, 2Department of Chemistry and Biotechnology, School of Engineering,The University of Tokyo, 3Faculty of Science and Engineering,Tecnologico de Monterrey, 4Jiksak Bioengineering, Inc.

简介：

Overview

This study presents a protocol for generating human iPS cell-derived motor nerve organoids through the spontaneous assembly of axons from a neuronal spheroid in a custom PDMS microculture chip. The platform allows for the investigation of axon bundle development and motor neuron diseases, facilitating efficient drug screening and testing.

Key Study Components

Area of Science

• Neuroscience
• Stem Cell Biology
• Organoid Technology

Background

• Motor nerve organoids provide a more physiological model compared to traditional in vitro systems.
• Understanding motor neuron diseases like Amyotrophic Lateral Sclerosis (ALS) is crucial for developing effective therapies.
• This protocol utilizes human iPS cells to generate a relevant biological model for research.

Purpose of Study

• To fabricate motor nerve organoids for studying axon development.
• To enhance drug screening for motor neuron diseases using a more accurate cellular environment.
• To provide a detailed methodology for generating and utilizing these models in research.

Methods Used

• The main platform used is a PDMS microfluidic chip designed for tissue culture.
• The biological model consists of motor neurons differentiated from human iPS cells.
• The protocol involves several key steps over a period of approximately two to three weeks.
• Cultures are maintained in a CO2 incubator, with specific media changes outlined for differentiation.
• Motor neuron spheroids are introduced into the microchannel to facilitate axon bundle formation.

Main Results

• Axons grow from motor neuron spheroids and assemble into organized bundles through axo-axonal interactions.
• Over 60% of differentiated cells express the motor neuron marker HB9, indicating successful differentiation.
• The model achieves functional maturation within 12 to 14 days post-differentiation, with key cellular changes noted over time.
• Motor nerve organoids show promise for biological analysis relevant to motor neuron diseases.

Conclusions

• This study demonstrates a reliable method for generating motor nerve organoids for research and pharmaceutical applications.
• It highlights the significance of using human iPS cells in disease modeling and drug screening.
• The findings contribute to a deeper understanding of neuronal mechanisms and the development of therapeutics.

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