Abstract
Human pluripotent stem cell (hPSC)-derived neurons provide exciting opportunities for in vitro modeling of neurological diseases and for advancing drug development and neurotoxicological studies. However, generating electrophysiologically mature neuronal networks from hPSCs has been challenging. Here, we report the differentiation of functionally active hPSC-derived cortical networks on defined laminin-521 substrate. We apply microelectrode array (MEA) measurements to assess network events and compare the activity development of hPSC-derived networks to that of widely used rat embryonic cortical cultures. In both of these networks, activity developed through a similar sequence of stages and time frames; however, the hPSC-derived networks showed unique patterns of bursting activity. The hPSC-derived networks developed synchronous activity, which involved glutamatergic and GABAergic inputs, recapitulating the classical cortical activity also observed in rodent counterparts. Principal component analysis (PCA) based on spike rates, network synchronization and burst features revealed the segregation of hPSC-derived and rat network recordings into different clusters, reflecting the species-specific and maturation state differences between the two networks. Overall, hPSC-derived neural cultures produced with a defined protocol generate cortical type network activity, which validates their applicability as a human-specific model for pharmacological studies and modeling network dysfunctions.
Highlights
Human pluripotent stem cell-derived neurons provide exciting opportunities for in vitro modeling of neurological diseases and for advancing drug development and neurotoxicological studies
The Human pluripotent stem cell (hPSC) were cultured in feeder-free conditions using LN521 substrate and Essential 8 (E8) medium according to the protocol from Hongisto et al (2017)
To assess the repeatability of the optimized culture protocol, experiments were performed with one human embryonic stem cell line (08/023) and two human induced pluripotent stem cell lines
Summary
Human pluripotent stem cell (hPSC)-derived neurons provide exciting opportunities for in vitro modeling of neurological diseases and for advancing drug development and neurotoxicological studies. We apply microelectrode array (MEA) measurements to assess network events and compare the activity development of hPSC-derived networks to that of widely used rat embryonic cortical cultures. HPSC-derived neural cultures produced with a defined protocol generate cortical type network activity, which validates their applicability as a human-specific model for pharmacological studies and modeling network dysfunctions. Human pluripotent stem cell (hPSC)-derived neurons possess great promise for unraveling the mechanisms of diseases with genetic susceptibility[1] They provide a human-specific model for in vitro research in neurotoxicology and drug discovery with the potential of reducing the use of animal studies[2]. We report that the differentiation of hPSC-derived cortical cultures on defined human recombinant laminin-521 (LN521) substrates is repeatable and results in the generation of cortical neurons, which form functionally active networks. The present data validate hPSC-derived neuronal networks as a reliable model for cortical activity development and consolidate their utilization in applications such as disease modeling, where network malfunctions are considered key characteristics
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