This experimental method has been validated in the eNeuro article "Neurons Derived from Human Induced Pluripotent Stem Cells Integrate into Rat Brain Circuits and Maintain Both Excitatory and Inhibitory Synaptic Activities." The original author published a detailed version of the method in the Bio-protocol journal, and everyone is welcome to enter the Bio-protocol journal to communicate directly with the author.
Introduction
In this Bio-protocol article, researchers from Johns Hopkins University School of Medicine in the United States describe a protocol for introducing human induced pluripotent stem cell (hiPSC)-derived neural precursor cells into rat brains.
Bright point
1. This method shows the coexistence of excitatory and inhibitory networks, enabling the development of a balanced network of excitatory and inhibitory neurons representing the complex structure of the human cortex.
2. This method provides a good in vivo model that more accurately represents the human cerebral cortex, allowing the study of interactions between excitatory and inhibitory networks under normal and pathological conditions.
3. The in vivo model constructed by this method can be used to study human cortical development and diseases related to imbalances of excitatory and inhibitory neurotransmission, such as autism, schizophrenia and Alzheimer's disease.
Background
Human neuronal transplantation offers new opportunities to mimic human neurological disorders and potential alternative therapies. However, the complex structure of the human cerebral cortex is divided into six layers, containing multiple neuronal subtypes in different cortexes (I-VI), which establish specific patterns of axonal output and dendritic input, provide the basic matrix of cortical circuits, with tightly interconnected excitatory and inhibitory neuronal networks, and the balance of excitatory and inhibitory neurotransmissions is necessary for normal brain function, which poses a major challenge to in vivo transplantation techniques to obtain balanced, functional, and homeostatically stable neuronal networks.
Human induced pluripotent stem cells (hiPSCs) allow the modeling of human neurological diseases in the context of human genetics. Currently, considerable progress has been made in establishing in vitro systems for differentiating hiPSCs into neurons, including the generation of excitatory (glutamatergic) projection neurons or inhibitory (GABAergic) interneurons. In vivo studies have also shown that hiPSC-derived neurons exhibit neuronal morphology and synaptic activity after transplantation into rodent brains. In this protocol, the authors present a protocol for introducing human induced pluripotent stem cell (hiPSC)-derived neural precursor cells into rat brains. Using this approach, hiPSC-derived neurons are structurally integrated into the rat forebrain, develop into functional neurons, exhibit electrophysiological properties, including firing, excitability, and inhibitory synaptic activity, and establish neuronal connections to host circuits.
Excerpt of the steps
step A. hiPSCs culture and passage
step B. Neural differentiation of hiPSCs
Figure 2. hiPSC-derived neurons show human cortical identity in culture after 10 weeks of differentiation.
step C. Animal transplantation
Figure 3. An outline showing transplantation of forebrain progenitors from hiPSCs (reproduced from Yin et al., 2019)
step D. Brain sections immunostaining
step E. Electrophysiological recordings of acute brain slices
Figure 5. hiPSC-derived neurons functionally integrate into the synaptic circuitry of the rat brain.
View the original article in the Bio-protocol journal for more information, including abstracts, background, consumables, reagents, instrument software, and more. Interested partners, copy the link below to have a look!
https://s.bio-protocol.org?s=190c36dddf71ea46
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Introduction to Bio-protocol
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