A neurosphere with human fetal brain derived neural stem cells differentiating into neurons

West Nile virus (WNV) and Usutu virus (USUV) are genetically related neurotropic mosquito-borne flaviviruses, which frequently co-circulate in nature. Despite USUV seeming to be less pathogenic for humans than WNV, the clinical manifestations induced by these two viruses often overlap and may evolve to produce severe neurological complications. The aim of this study was to investigate the effects of WNV and USUV infection on human induced pluripotent stem cell-derived neural stem cells (hNSCs), as a model of the neural progenitor cells in the developing fetal brain and in adult brain. Zika virus (ZIKV), a flavivirus with known tropism for NSCs, was used as the positive control. Infection of hNSCs and viral production, effects on cell viability, apoptosis, and innate antiviral responses were compared among viruses. WNV displayed the highest replication efficiency and cytopathic effects in hNSCs, followed by USUV and then ZIKV. In these cells, both WNV and USUV induced the overexpression of innate antiviral response genes at significantly higher levels than ZIKV. Expression of interferon type I, interleukin-1β and caspase-3 was significantly more elevated in WNV- than USUV-infected hNSCs, in agreement with the higher neuropathogenicity of WNV and the ability to inhibit the interferon response pathway.

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Functional maturation of human neural stem cells in a 3D bioengineered brain model enriched with fetal brain-derived matrix

10.1101/691907 ◽

2019 ◽

Cited By ~ 1

Author(s):

Disha Sood ◽

Dana M. Cairns ◽

Jayanth M. Dabbi ◽

Charu Ramakrishnan ◽

Karl Deisseroth ◽

...

Keyword(s):

Stem Cells ◽

Neural Stem Cells ◽

Brain Tissue ◽

Fetal Brain ◽

Functional Differentiation ◽

Adult Brain ◽

Human Neural Stem Cells ◽

Functional Maturation ◽

Enriched Cultures

AbstractBrain extracellular matrix (ECM) is often overlooked in vitro brain tissue models, despite its instructive roles during development. Using developmental stage-sourced brain ECM in reproducible 3D bioengineered culture systems, we demonstrate enhanced functional differentiation of human induced neural stem cells (hiNSCs) into healthy neurons and astrocytes. Particularly, fetal brain tissue-derived ECM supported long-term maintenance of differentiated neurons, demonstrated by morphology, gene expression and secretome profiling. Astrocytes were evident within the second month of differentiation, and reactive astrogliosis was inhibited in brain ECM-enriched cultures when compared to unsupplemented cultures. Functional maturation of the differentiated hiNSCs within fetal ECM-enriched cultures was confirmed by calcium signaling and unsupervised cluster analysis. Additionally, the study identified native biochemical cues in decellularized ECM with notable comparisons between fetal and adult brain-derived ECMs. The development of novel brain-specific biomaterials for generating mature in vitro brain models provides an important path forward for interrogation of neuron-glia interactions.

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Maternal sevoflurane exposure affects differentiation of hippocampal neural stem cells by regulating miR-410-3p and ATN1

10.21203/rs.3.rs-32498/v3 ◽

2020 ◽

Author(s):

Yi Zhang ◽

Ziyi Wu ◽

Xingyue Li ◽

Yuxiao Wan ◽

Yinong Zhang ◽

...

Keyword(s):

Stem Cells ◽

Neural Stem Cells ◽

Fetal Brain ◽

Repeated Exposure ◽

Luciferase Assay ◽

General Anesthetics ◽

Long Term Effects ◽

Pregnant Rats ◽

Early Differentiation

Abstract Background Currently, numerous animal studies have shown that exposure to commonly used general anesthetics during pregnancy may cause neurocognitive impairment in the offspring. Reportedly, exposure to sevoflurane during mid-trimester of pregnancy can inhibit proliferation of neural stem cells (NSCs) and lead to early apoptosis. Whether exposure to sevoflurane during pregnancy affects the differentiation of NSCs remains unclear. Methods In the present study, pregnant rats were exposed to 3% sevoflurane once for 2 h on gestational day 14 (G14) or 3 times for 2 h on G13, G14, and G15. Next, the differentiation of NSCs was measured using neuron marker β-tubulin III and astrocyte marker glial fibrillary acidic protein (GFAP) in fetal brain tissues 24 h and 72 h after anesthesia and in hippocampus on postnatal day 28. Primary cultured rat NSCs were exposed to 4.1% sevoflurane to explore the mechanism. Results The results showed that during mid-trimester, multiple exposures to sevoflurane can cause premature differentiation of NSCs in developing brains of offspring and lead to long-term neuron reduction and astrocyte proliferation in hippocampus. The data from the present study indicated that repeated exposure to sevoflurane downregulated atrophin-1 (ATN1) expression and caused early differentiation of NSCs. Overexpression of ATN1 via lentivirus transfection attenuated the influence of sevoflurane. Using dual luciferase assay, ATN1 was found to be a target gene of microRNA‐410-3p (miR‐410-3p). MiR-410-3p suppression via lentivirus transfection recovered the ATN1 expression and differentiation of NSCs.Conclusions The results from the present study demonstrated that repeated exposure to sevoflurane leads to early differentiation of NSCs and long-term effects via the miR-410-3p/ATN1 pathway.

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Maternal sevoflurane exposure affects differentiation of hippocampal neural stem cells by regulating miR-410-3p and ATN1

Stem Cell Research & Therapy ◽

10.1186/s13287-020-01936-9 ◽

2020 ◽

Vol 11 (1) ◽

Author(s):

Yi Zhang ◽

Ziyi Wu ◽

Xingyue Li ◽

Yuxiao Wan ◽

Yinong Zhang ◽

...

Keyword(s):

Stem Cells ◽

Neural Stem Cells ◽

Fetal Brain ◽

Repeated Exposure ◽

Luciferase Assay ◽

General Anesthetics ◽

Long Term Effects ◽

Pregnant Rats ◽

Early Differentiation

Abstract Background Currently, numerous animal studies have shown that exposure to commonly used general anesthetics during pregnancy may cause neurocognitive impairment in the offspring. Reportedly, exposure to sevoflurane during mid-trimester of pregnancy can inhibit proliferation of neural stem cells (NSCs) and lead to early apoptosis. Whether exposure to sevoflurane during pregnancy affects the differentiation of NSCs remains unclear. Methods In the present study, pregnant rats were exposed to 3% sevoflurane once for 2 h on gestational day 14 (G14) or 3 times for 2 h on G13, G14, and G15. Next, the differentiation of NSCs was measured using neuron marker β-tubulin III and astrocyte marker glial fibrillary acidic protein (GFAP) in fetal brain tissues 24 h and 72 h after anesthesia and in hippocampus on postnatal day 28. Primary cultured rat NSCs were exposed to 4.1% sevoflurane to explore the mechanism. Results The results showed that during mid-trimester, multiple exposures to sevoflurane can cause premature differentiation of NSCs in developing brains of offspring and lead to long-term neuron reduction and astrocyte proliferation in hippocampus. The data from the present study indicated that repeated exposure to sevoflurane downregulated atrophin-1 (ATN1) expression and caused early differentiation of NSCs. Overexpression of ATN1 via lentivirus transfection attenuated the influence of sevoflurane. Using dual luciferase assay, ATN1 was found to be a target gene of microRNA-410-3p (miR-410-3p). MiR-410-3p suppression via lentivirus transfection recovered the ATN1 expression and differentiation of NSCs. Conclusions The results from the present study demonstrated that repeated exposure to sevoflurane leads to early differentiation of NSCs and long-term effects via the miR-410-3p/ATN1 pathway.

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S185. IMPACT OF GENETIC LIABILITY TO SCHIZOPHRENIA ON IMPAIRED MIGRATION OF AUTOLOGOUS INDUCED NEURAL STEM CELLS

Schizophrenia Bulletin ◽

10.1093/schbul/sbaa031.251 ◽

2020 ◽

Vol 46 (Supplement_1) ◽

pp. S108-S109

Author(s):

Junhee Lee ◽

Sehyeon Song ◽

Tae Young Lee ◽

Mi-Sook Chang ◽

Jun Soo Kwon

Keyword(s):

Stem Cells ◽

Neural Stem Cells ◽

Fetal Brain ◽

Gene Signature ◽

University Hospital ◽

Mrna Levels ◽

Degree Relative ◽

Migration Assay ◽

Protein Levels ◽

Induced Neural Stem Cells

Abstract Background Since previous studies which presented the direct evidences of neurodevelopmental abnormality in schizophrenia using stem cell techniques, recent studies have demonstrated that induced neural stem cells (iNSCs) derived from patients with schizophrenia resembling their fetal brain tissue indeed shows aberrant migration and maintains the gene signature. However, the origin and nature of these abnormalities remain unresolved and no study examined unaffected individuals (UIs) with multiple relatives with schizophrenia. Methods Three patients diagnosed as schizophrenia (SZs), 3 UIs who has multiple relatives diagnosed as schizophrenia including at least one first-degree relative, and 3 healthy controls (HCs), all males, were recruited from Seoul National University Hospital (SNUH), Korea. iNSCs were generated from human adipose-derived stem cells (hADSCs) using small molecule-based lineage switch technique. The NSC marker expressions in iNSCs were analysed by real-time polymerase chain reaction (qPCR). The migration of iNSCs was analysed by CytoSelect 96-well cell migration assay. Results Demographic characteristics were not significantly different among the groups. Level of functioning was significantly lower in SZs. Either mRNA levels of NSC markers or NCAM protein levels were not significantly different among the groups. Doubling time among the groups were not significantly different. Migration capability of iNSCs from not only SZs, but also UIs were significantly smaller than HCs. Discussion We report a novel finding of defective ability of the iNSCs of UIs to reach the target along with those of SZs which is the first explicit demonstration of the postulation that neurodevelopmental abnormality might be present in UIs, using iNSCs directly derived from living subjects.

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Generation of spinal motor neurons from human fetal brain-derived neural stem cells: Role of basic fibroblast growth factor

Journal of Neuroscience Research ◽

10.1002/jnr.21856 ◽

2009 ◽

Vol 87 (2) ◽

pp. 318-332 ◽

Cited By ~ 34

Author(s):

Paivi M. Jordan ◽

Luis D. Ojeda ◽

Jason R. Thonhoff ◽

Junling Gao ◽

Darren Boehning ◽

...

Keyword(s):

Stem Cells ◽

Growth Factor ◽

Fibroblast Growth Factor ◽

Motor Neurons ◽

Fetal Brain ◽

Fibroblast Growth ◽

Human Fetal Brain ◽

Spinal Motor Neurons ◽

Spinal Motor

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Maternal sevoflurane exposure affects differentiation of hippocampal neural stem cells by regulating microRNA-410-3p and ATN1

10.21203/rs.3.rs-32498/v2 ◽

2020 ◽

Author(s):

YI ZHANG ◽

ZIYI WU ◽

XINGYUE LI ◽

YUXIAO WAN ◽

YINONG ZHANG ◽

...

Keyword(s):

Stem Cells ◽

Neural Stem Cells ◽

Fetal Brain ◽

Repeated Exposure ◽

Luciferase Assay ◽

General Anesthetics ◽

Long Term Effects ◽

Pregnant Rats ◽

Early Differentiation

Abstract Background Currently, numerous animal studies have shown that exposure to commonly used general anesthetics during pregnancy may cause neurocognitive impairment in the offspring. Reportedly, exposure to sevoflurane during mid-trimester of pregnancy can inhibit proliferation of neural stem cells (NSCs) and lead to early apoptosis. Whether exposure to sevoflurane during pregnancy affects the differentiation of NSCs remains unclear. Methods In the present study, pregnant rats were exposed to 3% sevoflurane once for 2 h on gestational day 14 (G14) or 3 times for 2 h on G13, G14, and G15. Next, the differentiation of NSCs was measured using neuron marker β-tubulin III and astrocyte marker glial fibrillary acidic protein (GFAP) in fetal brain tissues 24 h and 72 h after anesthesia and in hippocampus on postnatal day 28. Primary cultured rat NSCs were exposed to 4.1% sevoflurane to explore the mechanism. Results The results showed that during mid-trimester, multiple exposures to sevoflurane can cause premature differentiation of NSCs in developing brains of offspring and lead to long-term neuron reduction and astrocyte proliferation in hippocampus. The data from the present study indicated that repeated exposure to sevoflurane downregulated atrophin-1 (ATN1) expression and caused early differentiation of NSCs. Overexpression of ATN1 via lentivirus transfection attenuated the influence of sevoflurane. Using dual luciferase assay, ATN1 was found to be a target gene of microRNA‐410-3p (miR‐410-3p). MiR-410-3p suppression via lentivirus transfection recovered the ATN1 expression and differentiation of NSCs. Conclusions The results from the present study demonstrated that repeated exposure to sevoflurane leads to early differentiation of NSCs and long-term effects via the miR-410-3p/ATN1 pathway.

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