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a) Glutamatergic (E, in red) and GABAergic (I, in blue) neurons were derived from human induced pluripotent stem cells (hiPSCs) through the overexpression of Ngn2 and <t>Ascl1</t> transcriptional factors, respectively. b) The neurospheroids included 30% of rat astrocytes (in green) to favour the growth and the maturation of the neurons. The three experimental configurations included homogeneous excitatory (100E), heterogeneous (75E25I), and homogeneous inhibitory (100I) neurospheroids. c) Neurospheroids were generated by plating the cells in low-adhesion 96-wells to favour the self-organization into a three-dimensional structure and moved on high-density devices (3Brain GmbH) between DIV 63 and DIV 70 to record the electrophysiological activity. d) On top left: representative image of a neurospheroid plated on the active area of the high-density MEA; on top right: colour map in which each pixel represents one channel depicting the amplitude (in μV) of the electrophysiological activity of the same representative spheroid; on bottom left: extracellular 1-second signal trace of the same 3D network in which each box represents one channel; on bottom right: firing map of the same spheroid in which each pixel represents the mean firing rate of the channel.
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a) Glutamatergic (E, in red) and GABAergic (I, in blue) neurons were derived from human induced pluripotent stem cells (hiPSCs) through the overexpression of Ngn2 and <t>Ascl1</t> transcriptional factors, respectively. b) The neurospheroids included 30% of rat astrocytes (in green) to favour the growth and the maturation of the neurons. The three experimental configurations included homogeneous excitatory (100E), heterogeneous (75E25I), and homogeneous inhibitory (100I) neurospheroids. c) Neurospheroids were generated by plating the cells in low-adhesion 96-wells to favour the self-organization into a three-dimensional structure and moved on high-density devices (3Brain GmbH) between DIV 63 and DIV 70 to record the electrophysiological activity. d) On top left: representative image of a neurospheroid plated on the active area of the high-density MEA; on top right: colour map in which each pixel represents one channel depicting the amplitude (in μV) of the electrophysiological activity of the same representative spheroid; on bottom left: extracellular 1-second signal trace of the same 3D network in which each box represents one channel; on bottom right: firing map of the same spheroid in which each pixel represents the mean firing rate of the channel.
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a) Glutamatergic (E, in red) and GABAergic (I, in blue) neurons were derived from human induced pluripotent stem cells (hiPSCs) through the overexpression of Ngn2 and <t>Ascl1</t> transcriptional factors, respectively. b) The neurospheroids included 30% of rat astrocytes (in green) to favour the growth and the maturation of the neurons. The three experimental configurations included homogeneous excitatory (100E), heterogeneous (75E25I), and homogeneous inhibitory (100I) neurospheroids. c) Neurospheroids were generated by plating the cells in low-adhesion 96-wells to favour the self-organization into a three-dimensional structure and moved on high-density devices (3Brain GmbH) between DIV 63 and DIV 70 to record the electrophysiological activity. d) On top left: representative image of a neurospheroid plated on the active area of the high-density MEA; on top right: colour map in which each pixel represents one channel depicting the amplitude (in μV) of the electrophysiological activity of the same representative spheroid; on bottom left: extracellular 1-second signal trace of the same 3D network in which each box represents one channel; on bottom right: firing map of the same spheroid in which each pixel represents the mean firing rate of the channel.
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a) Glutamatergic (E, in red) and GABAergic (I, in blue) neurons were derived from human induced pluripotent stem cells (hiPSCs) through the overexpression of Ngn2 and <t>Ascl1</t> transcriptional factors, respectively. b) The neurospheroids included 30% of rat astrocytes (in green) to favour the growth and the maturation of the neurons. The three experimental configurations included homogeneous excitatory (100E), heterogeneous (75E25I), and homogeneous inhibitory (100I) neurospheroids. c) Neurospheroids were generated by plating the cells in low-adhesion 96-wells to favour the self-organization into a three-dimensional structure and moved on high-density devices (3Brain GmbH) between DIV 63 and DIV 70 to record the electrophysiological activity. d) On top left: representative image of a neurospheroid plated on the active area of the high-density MEA; on top right: colour map in which each pixel represents one channel depicting the amplitude (in μV) of the electrophysiological activity of the same representative spheroid; on bottom left: extracellular 1-second signal trace of the same 3D network in which each box represents one channel; on bottom right: firing map of the same spheroid in which each pixel represents the mean firing rate of the channel.
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a) Glutamatergic (E, in red) and GABAergic (I, in blue) neurons were derived from human induced pluripotent stem cells (hiPSCs) through the overexpression of Ngn2 and <t>Ascl1</t> transcriptional factors, respectively. b) The neurospheroids included 30% of rat astrocytes (in green) to favour the growth and the maturation of the neurons. The three experimental configurations included homogeneous excitatory (100E), heterogeneous (75E25I), and homogeneous inhibitory (100I) neurospheroids. c) Neurospheroids were generated by plating the cells in low-adhesion 96-wells to favour the self-organization into a three-dimensional structure and moved on high-density devices (3Brain GmbH) between DIV 63 and DIV 70 to record the electrophysiological activity. d) On top left: representative image of a neurospheroid plated on the active area of the high-density MEA; on top right: colour map in which each pixel represents one channel depicting the amplitude (in μV) of the electrophysiological activity of the same representative spheroid; on bottom left: extracellular 1-second signal trace of the same 3D network in which each box represents one channel; on bottom right: firing map of the same spheroid in which each pixel represents the mean firing rate of the channel.
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a) Glutamatergic (E, in red) and GABAergic (I, in blue) neurons were derived from human induced pluripotent stem cells (hiPSCs) through the overexpression of Ngn2 and <t>Ascl1</t> transcriptional factors, respectively. b) The neurospheroids included 30% of rat astrocytes (in green) to favour the growth and the maturation of the neurons. The three experimental configurations included homogeneous excitatory (100E), heterogeneous (75E25I), and homogeneous inhibitory (100I) neurospheroids. c) Neurospheroids were generated by plating the cells in low-adhesion 96-wells to favour the self-organization into a three-dimensional structure and moved on high-density devices (3Brain GmbH) between DIV 63 and DIV 70 to record the electrophysiological activity. d) On top left: representative image of a neurospheroid plated on the active area of the high-density MEA; on top right: colour map in which each pixel represents one channel depicting the amplitude (in μV) of the electrophysiological activity of the same representative spheroid; on bottom left: extracellular 1-second signal trace of the same 3D network in which each box represents one channel; on bottom right: firing map of the same spheroid in which each pixel represents the mean firing rate of the channel.
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a) Glutamatergic (E, in red) and GABAergic (I, in blue) neurons were derived from human induced pluripotent stem cells (hiPSCs) through the overexpression of Ngn2 and <t>Ascl1</t> transcriptional factors, respectively. b) The neurospheroids included 30% of rat astrocytes (in green) to favour the growth and the maturation of the neurons. The three experimental configurations included homogeneous excitatory (100E), heterogeneous (75E25I), and homogeneous inhibitory (100I) neurospheroids. c) Neurospheroids were generated by plating the cells in low-adhesion 96-wells to favour the self-organization into a three-dimensional structure and moved on high-density devices (3Brain GmbH) between DIV 63 and DIV 70 to record the electrophysiological activity. d) On top left: representative image of a neurospheroid plated on the active area of the high-density MEA; on top right: colour map in which each pixel represents one channel depicting the amplitude (in μV) of the electrophysiological activity of the same representative spheroid; on bottom left: extracellular 1-second signal trace of the same 3D network in which each box represents one channel; on bottom right: firing map of the same spheroid in which each pixel represents the mean firing rate of the channel.
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a) Glutamatergic (E, in red) and GABAergic (I, in blue) neurons were derived from human induced pluripotent stem cells (hiPSCs) through the overexpression of Ngn2 and <t>Ascl1</t> transcriptional factors, respectively. b) The neurospheroids included 30% of rat astrocytes (in green) to favour the growth and the maturation of the neurons. The three experimental configurations included homogeneous excitatory (100E), heterogeneous (75E25I), and homogeneous inhibitory (100I) neurospheroids. c) Neurospheroids were generated by plating the cells in low-adhesion 96-wells to favour the self-organization into a three-dimensional structure and moved on high-density devices (3Brain GmbH) between DIV 63 and DIV 70 to record the electrophysiological activity. d) On top left: representative image of a neurospheroid plated on the active area of the high-density MEA; on top right: colour map in which each pixel represents one channel depicting the amplitude (in μV) of the electrophysiological activity of the same representative spheroid; on bottom left: extracellular 1-second signal trace of the same 3D network in which each box represents one channel; on bottom right: firing map of the same spheroid in which each pixel represents the mean firing rate of the channel.
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Image Search Results


a) Glutamatergic (E, in red) and GABAergic (I, in blue) neurons were derived from human induced pluripotent stem cells (hiPSCs) through the overexpression of Ngn2 and Ascl1 transcriptional factors, respectively. b) The neurospheroids included 30% of rat astrocytes (in green) to favour the growth and the maturation of the neurons. The three experimental configurations included homogeneous excitatory (100E), heterogeneous (75E25I), and homogeneous inhibitory (100I) neurospheroids. c) Neurospheroids were generated by plating the cells in low-adhesion 96-wells to favour the self-organization into a three-dimensional structure and moved on high-density devices (3Brain GmbH) between DIV 63 and DIV 70 to record the electrophysiological activity. d) On top left: representative image of a neurospheroid plated on the active area of the high-density MEA; on top right: colour map in which each pixel represents one channel depicting the amplitude (in μV) of the electrophysiological activity of the same representative spheroid; on bottom left: extracellular 1-second signal trace of the same 3D network in which each box represents one channel; on bottom right: firing map of the same spheroid in which each pixel represents the mean firing rate of the channel.

Journal: bioRxiv

Article Title: Functional human neurospheroids recapitulate key features of cortical complexity

doi: 10.64898/2026.03.09.710475

Figure Lengend Snippet: a) Glutamatergic (E, in red) and GABAergic (I, in blue) neurons were derived from human induced pluripotent stem cells (hiPSCs) through the overexpression of Ngn2 and Ascl1 transcriptional factors, respectively. b) The neurospheroids included 30% of rat astrocytes (in green) to favour the growth and the maturation of the neurons. The three experimental configurations included homogeneous excitatory (100E), heterogeneous (75E25I), and homogeneous inhibitory (100I) neurospheroids. c) Neurospheroids were generated by plating the cells in low-adhesion 96-wells to favour the self-organization into a three-dimensional structure and moved on high-density devices (3Brain GmbH) between DIV 63 and DIV 70 to record the electrophysiological activity. d) On top left: representative image of a neurospheroid plated on the active area of the high-density MEA; on top right: colour map in which each pixel represents one channel depicting the amplitude (in μV) of the electrophysiological activity of the same representative spheroid; on bottom left: extracellular 1-second signal trace of the same 3D network in which each box represents one channel; on bottom right: firing map of the same spheroid in which each pixel represents the mean firing rate of the channel.

Article Snippet: GABAergic neurons were derived from C2 by overexpressing mouse neuronal determinant Ascl1 (Addgene, 97329) upon doxycycline treatment and supplementation with forskolin (10 μM, Sigma Aldrich) .

Techniques: Derivative Assay, Over Expression, Generated, Activity Assay