Spinal dI4 Interneuron Differentiation From Human Pluripotent Stem Cells

Frontiers in Molecular Neuroscience, 2022 · DOI: 10.3389/fnmol.2022.845875 · Published: April 8, 2022

Spinal Cord Injury Neurology Regenerative Medicine & Stem Cells

Simple Explanation

Spinal interneurons (INs) are crucial for relaying nerve signals and regulating motor functions. This study focuses on creating spinal dI4 inhibitory GABAergic INs from human pluripotent stem cells (hPSCs). The protocol involves guiding hPSCs through neuroepithelial induction, spinal progenitor patterning, progenitor expansion, and finally, neuronal differentiation, using specific molecules to direct cell fate. The ability to generate enriched spinal dI4 GABAergicINs will likely facilitate the study of human spinal IN development and regenerative therapies for traumatic injuries and diseases of the spinal cord.

Study Duration

Not specified

Participants

Human Pluripotent Stem Cells

Evidence Level

Not specified

Key Findings

1
A reproducible protocol was devised to differentiate human pluripotent stem cells (hPSCs) from enriched spinal dI4 inhibitory GABAergic INs.
2
The protocol employs morphogen activators and inhibitors like retinoic acid (RA) and cyclopamine (CYC) to generate highly enriched dI4 progenitors.
3
The resulting dI4 neurons express key markers such as Pax2, Lhx1/5, GABA, and NF-200, indicating successful differentiation into spinal dI4 GABAergic INs.

Research Summary

This study presents a detailed protocol for differentiating human pluripotent stem cells (hPSCs) into spinal dI4 GABAergic interneurons (INs), which are crucial for spinal cord function and potential therapeutic targets. The differentiation process involves four stages: neuroepithelial induction, spinal progenitor patterning using morphogens (RA and CYC), progenitor expansion through suspension culture, and final differentiation into mature dI4 INs. The generated dI4 INs exhibit appropriate markers and characteristics, resembling those in the developing human spinal cord, making them valuable for studying spinal cord development and potential regenerative therapies.

Practical Implications

Therapeutic Potential

The generated dI4 GABAergic INs can be transplanted into the spinal cord to explore their integration and therapeutic potential in treating spinal cord injuries and diseases.

Drug Screening Platforms

The protocol enables the generation of engineered spinal INs from hPSCs to establish platforms for screening or testing compounds that regulate IN functions.

Understanding Spinal Cord Development

The protocol provides a valuable tool for studying human spinal IN subtype development, offering insights into the cellular and molecular mechanisms involved.

Study Limitations

1
The study does not specify the exact duration of the study.
2
The study does not detail the long-term functional integration of the derived interneurons in vivo.
3
The protocol's efficiency may vary depending on the specific hPSC line used.

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