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IPSC-Derived Human Neurons with GCaMP6s Expression Allow In Vitro Study of Neurophysiological Responses to
A A Galiakberova1,2, A M Surin3,4, Z V Bakaeva3,5
1Center for Precision Genome Editing and Genetic Technologies for Biomedicine, Pirogov Russian National Research Medical University, Ostrovitianov Street, Moscow, Russia, 117997. adgaliakberova@gmail.com.
Neurochemical Research
|December 2, 2021
Summary
Researchers developed a new method to create human neurons in vitro using human induced pluripotent stem cells (hiPSCs). This model effectively studies neurochemical effects on neuronal activity via calcium imaging.
Area of Science:
- Neuroscience
- Stem Cell Biology
- Biotechnology
Background:
- Studying human neurons and neurochemicals is challenging due to limited primary biomaterial access.
- Advances in stem cell technology enable the creation of in vitro human neuron models.
Purpose of the Study:
- To develop an efficient method for generating human neurons expressing the GCaMP6s calcium indicator.
- To establish a robust in vitro model for studying neurochemical interactions and neurophysiology.
Main Methods:
- Utilized a human induced pluripotent stem cell (hiPSC) line with a TetON-NGN2 transgene.
- Developed a protocol for neuronal differentiation and GCaMP6s indicator expression.
- Employed fluorescence microscopy for registering neurophysiological activity.
Main Results:
- Successfully generated substantial quantities of human neurons suitable for research.
- Demonstrated that glutamate (Glu) induces intracellular calcium ([Ca2+]i) increases via ionotropic receptors.
- Identified predominant involvement of NMDA-type receptors in the glutamate response.
Conclusions:
- The developed method provides a valuable and efficient tool for in vitro neuroscience research.
- The human iPSC-derived neuron model successfully captures neurochemical-induced activity.
- This technology facilitates the study of neurochemical effects on specific neurophysiological responses.
