Fully defined NGN2 neuron protocol reveals diverse signatures of neuronal maturation

Xiwei Shan; Ai Zhang; Mitchell G. Rezzonico; Ming Chi Tsai; Carlos Sanchez-Priego; Yingjie Zhang; Michelle B. Chen; Meena Choi; José Miguel Andrade López; Lilian Phu; Amber L. Cramer; Qiao Zhang; Jillian M. Pattison; Christopher M. Rose; Casper C. Hoogenraad; Claire G. Jeong

doi:10.1016/j.crmeth.2024.100858

Fully defined NGN2 neuron protocol reveals diverse signatures of neuronal maturation

Xiwei Shan, Ai Zhang, Mitchell G. Rezzonico, Ming Chi Tsai, Carlos Sanchez-Priego, Yingjie Zhang, Michelle B. Chen, Meena Choi, José Miguel Andrade López, Lilian Phu, Amber L. Cramer, Qiao Zhang, Jillian M. Pattison, Christopher M. Rose, Casper C. Hoogenraad, Claire G. Jeong

Research output: Contribution to journal › Article › peer-review

Abstract

NGN2-driven induced pluripotent stem cell (iPSC)-to-neuron conversion is a popular method for human neurological disease modeling. In this study, we present a standardized approach for generating neurons utilizing clonal, targeted-engineered iPSC lines with defined reagents. We demonstrate consistent production of excitatory neurons at scale and long-term maintenance for at least 150 days. Temporal omics, electrophysiological, and morphological profiling indicate continued maturation to postnatal-like neurons. Quantitative characterizations through transcriptomic, imaging, and functional assays reveal coordinated actions of multiple pathways that drive neuronal maturation. We also show the expression of disease-related genes in these neurons to demonstrate the relevance of our protocol for modeling neurological disorders. Finally, we demonstrate efficient generation of NGN2-integrated iPSC lines. These workflows, profiling data, and functional characterizations enable the development of reproducible human in vitro models of neurological disorders.

Original language	English
Article number	100858
Journal	Cell Reports Methods
Volume	4
Issue number	9
DOIs	https://doi.org/10.1016/j.crmeth.2024.100858
State	Published - 16 Sep 2024
Externally published	Yes

Keywords

CP: neuroscience
CP: stem cell
◼

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10.1016/j.crmeth.2024.100858

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Shan, X., Zhang, A., Rezzonico, M. G., Tsai, M. C., Sanchez-Priego, C., Zhang, Y., Chen, M. B., Choi, M., Andrade López, J. M., Phu, L., Cramer, A. L., Zhang, Q., Pattison, J. M., Rose, C. M., Hoogenraad, C. C., & Jeong, C. G. (2024). Fully defined NGN2 neuron protocol reveals diverse signatures of neuronal maturation. Cell Reports Methods, 4(9), Article 100858. https://doi.org/10.1016/j.crmeth.2024.100858

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title = "Fully defined NGN2 neuron protocol reveals diverse signatures of neuronal maturation",

abstract = "NGN2-driven induced pluripotent stem cell (iPSC)-to-neuron conversion is a popular method for human neurological disease modeling. In this study, we present a standardized approach for generating neurons utilizing clonal, targeted-engineered iPSC lines with defined reagents. We demonstrate consistent production of excitatory neurons at scale and long-term maintenance for at least 150 days. Temporal omics, electrophysiological, and morphological profiling indicate continued maturation to postnatal-like neurons. Quantitative characterizations through transcriptomic, imaging, and functional assays reveal coordinated actions of multiple pathways that drive neuronal maturation. We also show the expression of disease-related genes in these neurons to demonstrate the relevance of our protocol for modeling neurological disorders. Finally, we demonstrate efficient generation of NGN2-integrated iPSC lines. These workflows, profiling data, and functional characterizations enable the development of reproducible human in vitro models of neurological disorders.",

keywords = "CP: neuroscience, CP: stem cell, ◼",

author = "Xiwei Shan and Ai Zhang and Rezzonico, {Mitchell G.} and Tsai, {Ming Chi} and Carlos Sanchez-Priego and Yingjie Zhang and Chen, {Michelle B.} and Meena Choi and {Andrade L{\'o}pez}, {Jos{\'e} Miguel} and Lilian Phu and Cramer, {Amber L.} and Qiao Zhang and Pattison, {Jillian M.} and Rose, {Christopher M.} and Hoogenraad, {Casper C.} and Jeong, {Claire G.}",

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year = "2024",

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day = "16",

doi = "10.1016/j.crmeth.2024.100858",

language = "English",

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Shan, X, Zhang, A, Rezzonico, MG, Tsai, MC, Sanchez-Priego, C, Zhang, Y, Chen, MB, Choi, M, Andrade López, JM, Phu, L, Cramer, AL, Zhang, Q, Pattison, JM, Rose, CM, Hoogenraad, CC & Jeong, CG 2024, 'Fully defined NGN2 neuron protocol reveals diverse signatures of neuronal maturation', Cell Reports Methods, vol. 4, no. 9, 100858. https://doi.org/10.1016/j.crmeth.2024.100858

TY - JOUR

T1 - Fully defined NGN2 neuron protocol reveals diverse signatures of neuronal maturation

AU - Shan, Xiwei

AU - Zhang, Ai

AU - Rezzonico, Mitchell G.

AU - Tsai, Ming Chi

AU - Sanchez-Priego, Carlos

AU - Zhang, Yingjie

AU - Chen, Michelle B.

AU - Choi, Meena

AU - Andrade López, José Miguel

AU - Phu, Lilian

AU - Cramer, Amber L.

AU - Zhang, Qiao

AU - Pattison, Jillian M.

AU - Rose, Christopher M.

AU - Hoogenraad, Casper C.

AU - Jeong, Claire G.

PY - 2024/9/16

Y1 - 2024/9/16

N2 - NGN2-driven induced pluripotent stem cell (iPSC)-to-neuron conversion is a popular method for human neurological disease modeling. In this study, we present a standardized approach for generating neurons utilizing clonal, targeted-engineered iPSC lines with defined reagents. We demonstrate consistent production of excitatory neurons at scale and long-term maintenance for at least 150 days. Temporal omics, electrophysiological, and morphological profiling indicate continued maturation to postnatal-like neurons. Quantitative characterizations through transcriptomic, imaging, and functional assays reveal coordinated actions of multiple pathways that drive neuronal maturation. We also show the expression of disease-related genes in these neurons to demonstrate the relevance of our protocol for modeling neurological disorders. Finally, we demonstrate efficient generation of NGN2-integrated iPSC lines. These workflows, profiling data, and functional characterizations enable the development of reproducible human in vitro models of neurological disorders.

AB - NGN2-driven induced pluripotent stem cell (iPSC)-to-neuron conversion is a popular method for human neurological disease modeling. In this study, we present a standardized approach for generating neurons utilizing clonal, targeted-engineered iPSC lines with defined reagents. We demonstrate consistent production of excitatory neurons at scale and long-term maintenance for at least 150 days. Temporal omics, electrophysiological, and morphological profiling indicate continued maturation to postnatal-like neurons. Quantitative characterizations through transcriptomic, imaging, and functional assays reveal coordinated actions of multiple pathways that drive neuronal maturation. We also show the expression of disease-related genes in these neurons to demonstrate the relevance of our protocol for modeling neurological disorders. Finally, we demonstrate efficient generation of NGN2-integrated iPSC lines. These workflows, profiling data, and functional characterizations enable the development of reproducible human in vitro models of neurological disorders.

KW - CP: neuroscience

KW - CP: stem cell

KW - ◼

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DO - 10.1016/j.crmeth.2024.100858

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SN - 2667-2375

VL - 4

JO - Cell Reports Methods

JF - Cell Reports Methods

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ER -

Fully defined NGN2 neuron protocol reveals diverse signatures of neuronal maturation

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Keywords

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