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Human cortical neurons rapidly generated by embryonic stem cell programming integrate into the stroke-injured rat cortex

Martinez-Curiel, Raquel LU orcid ; Hajy, Mazin LU ; Tsupykov, Oleg ; Jansson, Linda LU ; Avaliani, Natalia LU ; Tampé, Juliane LU orcid ; Monni, Emanuela LU ; Skibo, Galyna ; Lindvall, Olle LU and Palma-Tortosa, Sara LU , et al. (2025) In Stem Cells 43(11).
Abstract

Stem cell sources capable of producing appropriate cells for replacement will be necessary for functional repair of the injured brain. Here, we have determined whether transcription factor programming of human embryonic stem (hES) cells can be used to generate layer-specific cortical neurons capable of integrating into the stroke-injured rat cortex. Human embryonic stem cells were programmed via overexpression of neurogenin 2 (NGN2). After 7days, hES-induced neurons (hES-iNs) were characterized in vitro using immunocytochemistry, RT-qPCR, and whole-cell patch-clamp. Cortical ischemic stroke was induced in rats via distal middle cerebral artery occlusion. Forty-eight hours later, hES-iNs were transplanted into the somatosensory cortex... (More)

Stem cell sources capable of producing appropriate cells for replacement will be necessary for functional repair of the injured brain. Here, we have determined whether transcription factor programming of human embryonic stem (hES) cells can be used to generate layer-specific cortical neurons capable of integrating into the stroke-injured rat cortex. Human embryonic stem cells were programmed via overexpression of neurogenin 2 (NGN2). After 7days, hES-induced neurons (hES-iNs) were characterized in vitro using immunocytochemistry, RT-qPCR, and whole-cell patch-clamp. Cortical ischemic stroke was induced in rats via distal middle cerebral artery occlusion. Forty-eight hours later, hES-iNs were transplanted into the somatosensory cortex adjacent to the ischemic lesion. Three months thereafter, brains were analyzed for expression of neuronal markers, axonal myelination, and synapse formation using immunohistochemistry and immunoelectron microscopy (iEM). Overexpression of NGN2 in hES cells for 7days generated excitatory neurons, expressing cortical markers at different stages of maturation. After transplantation, the hES-iNs expressed markers of both immature and mature neurons and of upper and deep cortical layers. The hES-iNs sent widespread projections to both hemispheres, and iEM revealed that they were myelinated by host oligodendrocytes and had formed efferent synaptic connections with host cortical neurons. The hES cells programmed via NGN2 overexpression gave rise to subtypes of cortical neurons, capable of integrating structurally into the injured brain, more rapidly than neurons produced by previous protocols. Functional characterization of the grafted hES-iNs and their impact on the balance between brain excitation and inhibition are now highly warranted. This new stem cell source should be considered when, in the future, the most suitable candidate will be selected for clinical translation.

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organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
cell transplantation, human embryonic stem cells, induced neurons, ischemic stroke, restoration
in
Stem Cells
volume
43
issue
11
article number
sxaf049
publisher
Oxford University Press
external identifiers
  • pmid:40650995
  • scopus:105019078339
ISSN
1066-5099
DOI
10.1093/stmcls/sxaf049
language
English
LU publication?
yes
additional info
Publisher Copyright: © The Author(s) 2025. Published by Oxford University Press.
id
b3547abe-b0c3-4ca0-b4b0-44f4277088f2
date added to LUP
2025-12-16 15:48:56
date last changed
2025-12-17 03:03:47
@article{b3547abe-b0c3-4ca0-b4b0-44f4277088f2,
  abstract     = {{<p>Stem cell sources capable of producing appropriate cells for replacement will be necessary for functional repair of the injured brain. Here, we have determined whether transcription factor programming of human embryonic stem (hES) cells can be used to generate layer-specific cortical neurons capable of integrating into the stroke-injured rat cortex. Human embryonic stem cells were programmed via overexpression of neurogenin 2 (NGN2). After 7days, hES-induced neurons (hES-iNs) were characterized in vitro using immunocytochemistry, RT-qPCR, and whole-cell patch-clamp. Cortical ischemic stroke was induced in rats via distal middle cerebral artery occlusion. Forty-eight hours later, hES-iNs were transplanted into the somatosensory cortex adjacent to the ischemic lesion. Three months thereafter, brains were analyzed for expression of neuronal markers, axonal myelination, and synapse formation using immunohistochemistry and immunoelectron microscopy (iEM). Overexpression of NGN2 in hES cells for 7days generated excitatory neurons, expressing cortical markers at different stages of maturation. After transplantation, the hES-iNs expressed markers of both immature and mature neurons and of upper and deep cortical layers. The hES-iNs sent widespread projections to both hemispheres, and iEM revealed that they were myelinated by host oligodendrocytes and had formed efferent synaptic connections with host cortical neurons. The hES cells programmed via NGN2 overexpression gave rise to subtypes of cortical neurons, capable of integrating structurally into the injured brain, more rapidly than neurons produced by previous protocols. Functional characterization of the grafted hES-iNs and their impact on the balance between brain excitation and inhibition are now highly warranted. This new stem cell source should be considered when, in the future, the most suitable candidate will be selected for clinical translation.</p>}},
  author       = {{Martinez-Curiel, Raquel and Hajy, Mazin and Tsupykov, Oleg and Jansson, Linda and Avaliani, Natalia and Tampé, Juliane and Monni, Emanuela and Skibo, Galyna and Lindvall, Olle and Palma-Tortosa, Sara and Kokaia, Zaal}},
  issn         = {{1066-5099}},
  keywords     = {{cell transplantation; human embryonic stem cells; induced neurons; ischemic stroke; restoration}},
  language     = {{eng}},
  month        = {{11}},
  number       = {{11}},
  publisher    = {{Oxford University Press}},
  series       = {{Stem Cells}},
  title        = {{Human cortical neurons rapidly generated by embryonic stem cell programming integrate into the stroke-injured rat cortex}},
  url          = {{http://dx.doi.org/10.1093/stmcls/sxaf049}},
  doi          = {{10.1093/stmcls/sxaf049}},
  volume       = {{43}},
  year         = {{2025}},
}