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Differentiation of Mouse Embryonic Stem Cells into Cortical Interneuron Precursors
Published on: December 3, 2017
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Generation of v2a interneurons from mouse embryonic stem cells
Chelsea R Brown1, Jessica C Butts, Dylan A McCreedy
1Department of Biomedical Engineering, Washington University in St. Louis , St. Louis, Missouri.
Stem Cells and Development
|March 22, 2014
Summary
Researchers developed a new method to create V2a interneurons from mouse stem cells. This protocol enhances V2a interneuron differentiation for crucial research in neurobiology and motor control.
Area of Science:
- Neuroscience
- Developmental Biology
- Stem Cell Research
Background:
- V2a interneurons are vital for central pattern generators (CPGs) controlling locomotion, skilled reaching, and respiration.
- Current in vitro study limitations exist due to restricted sources of V2a interneurons.
Purpose of the Study:
- To establish a differentiation protocol for generating V2a interneurons from mouse embryonic stem cells (mESCs).
- To optimize conditions for V2a interneuron production for research applications.
Main Methods:
- Utilized a 2(-)/4(+) induction protocol with varying retinoic acid (RA) and purmorphamine (Pur) concentrations.
- Inhibited Notch signaling to promote V2a over V2b interneuron commitment.
- Assessed cell commitment via qRT-PCR, immunocytochemistry, and flow cytometry for specific transcription factors.
Main Results:
- Low RA and high Pur concentrations increased V2a interneuron-specific transcription factors.
- Notch inhibition significantly favored V2a interneuron differentiation compared to V2b.
- The protocol successfully generated V2a interneurons from mESCs.
Conclusions:
- The developed protocol provides a reliable method for producing V2a interneurons in vitro.
- This advancement supports further research into V2a interneuron development and function.
- Enables new avenues for developmental neurobiology, electrophysiology, and transplantation studies.
