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Author Spotlight: Hypothalamic Neural Mechanism Insights
Published on: August 4, 2023
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Differentiation, Transcriptomic Profiling, and Calcium Imaging of Human Hypothalamic Neurons
Hsiao-Jou Cortina Chen1,2, Simone Mazzaferro1,2, Tian Tian1,2
1Metabolic Research Laboratories, Wellcome Trust MRC Institute of Metabolic Science, University of Cambridge, Addenbrooke's Hospital, Cambridge, UK.
Current Protocols
|June 5, 2023
Summary
This study provides updated protocols for differentiating human pluripotent stem cells (hPSCs) into hypothalamic neurons. These methods enable research into neurological disorders like obesity and diabetes.
Area of Science:
- Neuroscience
- Stem Cell Biology
- Developmental Biology
Background:
- The hypothalamus regulates vital homeostatic processes and behaviors.
- Human pluripotent stem cells (hPSCs) offer a model for generating hypothalamic cell types.
- Existing protocols require optimization for broader accessibility and application.
Purpose of the Study:
- To present updated protocols for differentiating and maturing human hypothalamic neurons from hPSCs.
- To incorporate advanced techniques for analyzing these neuronal populations.
- To facilitate the use of this model system for disease modeling and therapeutic translation.
Main Methods:
- hPSC maintenance and hypothalamic neuron differentiation.
- Neuronal maturation, cryopreservation, and thawing of progenitors.
- Transcriptomic profiling (Bulk RNA sequencing) and calcium imaging (Fura-2 AM, Rhod-3 AM).
Main Results:
- Established protocols for generating and maturing hypothalamic neurons and progenitors.
- Included methods for quality control, cryopreservation, and advanced cellular analysis.
- Demonstrated utility for studying hypothalamic function and disease.
Conclusions:
- These comprehensive protocols enhance the utility of hPSC-derived hypothalamic neurons for fundamental research.
- The updated methods support the investigation of diseases including obesity, diabetes, and stress disorders.
- This model system is valuable for both basic biological discovery and therapeutic development.
