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Human Pluripotent Stem Cell-Derived Astrocyte Functionality Compares Favorably with Primary Rat Astrocytes
Bas Lendemeijer1,2,3, Maurits Unkel1, Hilde Smeenk1
1Department of Psychiatry, Erasmus University Medical Center, Rotterdam 3015 AA, The Netherlands.
Eneuro
|September 3, 2024
Summary
Human pluripotent stem cell-derived astrocytes support neural network activity comparable to rodent astrocytes. This provides a fully human model for studying astrocyte function and neuronal-glial interactions in neurological research.
Area of Science:
- Neuroscience
- Stem Cell Biology
- Astrocyte Biology
Background:
- Astrocytes are crucial for neural network formation and maintenance.
- Obtaining functional human astrocytes for research has been a significant challenge.
- Primary rodent astrocytes are currently the gold standard in co-culture models with human neurons.
Purpose of the Study:
- To develop a method for generating pure, functional human astrocytes from human pluripotent stem cells (hPSCs).
- To compare the ability of hPSC-derived astrocytes to support human neural network activity and maturation against primary rat astrocytes.
- To establish a fully human in vitro model for studying astrocyte function and neuronal-glial interactions.
Main Methods:
- Directed differentiation of hPSC-derived neural precursor cells into astroglia using leukemia inhibitory factor (LIF) and bone morphogenetic protein-4 (BMP4).
- Single-cell RNA sequencing to confirm astroglial identity and assess transcriptional adaptations.
- Co-culture of hPSC-derived astrocytes with human neurons.
- Multielectrode array (MEA) recordings to measure network activity.
- Whole-cell patch-clamp recordings to analyze synaptic currents.
- Synapse density quantification.
Main Results:
- A combination of LIF and BMP4 efficiently generated a highly pure population of astroglia from hPSCs within 28 days.
- hPSC-derived astrocytes supported robust human neural network activity, comparable to rat astrocytes in terms of network frequency.
- Co-culture with hPSC-derived astrocytes significantly increased human neuron spike frequency, postsynaptic currents, and synapse density compared to rat astrocytes.
- Transcriptional analysis revealed maturation of both hPSC-derived astrocytes and neurons in co-culture.
Conclusions:
- hPSC-derived astrocytes are a viable and effective alternative to primary rodent astrocytes for supporting human neural network activity and maturation.
- This study provides a scalable, fully human in vitro platform for investigating astrocyte biology and its role in neurological diseases.
- The findings pave the way for more accurate modeling of human neuronal-glial interactions and the development of novel therapeutic strategies.
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