Functional Expression of Choline Transporters in Human Neural Stem Cells and Its Link to Cell Proliferation, Cell Viability, and Neurite Outgrowth

Choline and choline metabolites are essential for all cellular functions. They have also been reported to be crucial for neural development. In this work, we studied the functional characteristics of the choline uptake system in human neural stem cells (hNSCs). Additionally, we investigated the effect of extracellular choline uptake inhibition on the cellular activities in hNSCs. We found that the mRNAs and proteins of choline transporter-like protein 1 (CTL1) and CTL2 were expressed at high levels. Immunostaining showed that CTL1 and CTL2 were localized in the cell membrane and partly in the mitochondria, respectively. The uptake of extracellular choline was saturable and performed by a single uptake mechanism, which was Na+-independent and pH-dependent. We conclude that CTL1 is responsible for extracellular choline uptake, and CTL2 may uptake choline in the mitochondria and be involved in DNA methylation via choline oxidation. Extracellular choline uptake inhibition caused intracellular choline deficiency in hNSCs, which suppressed cell proliferation, cell viability, and neurite outgrowth. Our findings contribute to the understanding of the role of choline in neural development as well as the pathogenesis of various neurological diseases caused by choline deficiency or choline uptake impairment.

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Functional Expression of Choline Transporters in Human Neural Stem Cells and Its Link to Cell Proliferation, Cell Viability, and Neurite Outgrowth

10.20944/preprints202102.0009.v1 ◽

2021 ◽

Author(s):

Yosuke Fujita ◽

Tomoki Nagakura ◽

Hiroyuki Uchino ◽

Masato Inazu ◽

Tsuyoshi Yamanaka

Keyword(s):

Stem Cells ◽

Cell Proliferation ◽

Neural Stem Cells ◽

Cell Viability ◽

Neurite Outgrowth ◽

Neural Development ◽

Choline Uptake ◽

Choline Deficiency ◽

Uptake Inhibition ◽

Human Neural Stem Cells

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Icariin promotes cell proliferation and regulates gene expression in human neural stem cells in vitro

Molecular Medicine Reports ◽

10.3892/mmr.2016.5377 ◽

2016 ◽

Vol 14 (2) ◽

pp. 1316-1322 ◽

Cited By ~ 9

Author(s):

Pan Yang ◽

Yun-Qian Guan ◽

Ya-Li Li ◽

Li Zhang ◽

Lan Zhang ◽

...

Keyword(s):

Gene Expression ◽

Stem Cells ◽

Cell Proliferation ◽

Neural Stem Cells ◽

Human Neural Stem Cells

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mRNA-Driven Generation of Transgene-Free Neural Stem Cells from Human Urine-Derived Cells

Cells ◽

10.3390/cells8091043 ◽

2019 ◽

Vol 8 (9) ◽

pp. 1043 ◽

Cited By ~ 1

Author(s):

Phil Jun Kang ◽

Daryeon Son ◽

Tae Hee Ko ◽

Wonjun Hong ◽

Wonjin Yun ◽

...

Keyword(s):

Stem Cells ◽

Neural Stem Cells ◽

Human Urine ◽

Neurological Disorders ◽

Therapeutic Potential ◽

Embryonic Stem ◽

Global Gene Expression ◽

Patient Specific ◽

Human Neural Stem Cells

Human neural stem cells (NSCs) hold enormous promise for neurological disorders, typically requiring their expandable and differentiable properties for regeneration of damaged neural tissues. Despite the therapeutic potential of induced NSCs (iNSCs), a major challenge for clinical feasibility is the presence of integrated transgenes in the host genome, contributing to the risk for undesired genotoxicity and tumorigenesis. Here, we describe the advanced transgene-free generation of iNSCs from human urine-derived cells (HUCs) by combining a cocktail of defined small molecules with self-replicable mRNA delivery. The established iNSCs were completely transgene-free in their cytosol and genome and further resembled human embryonic stem cell-derived NSCs in the morphology, biological characteristics, global gene expression, and potential to differentiate into functional neurons, astrocytes, and oligodendrocytes. Moreover, iNSC colonies were observed within eight days under optimized conditions, and no teratomas formed in vivo, implying the absence of pluripotent cells. This study proposes an approach to generate transplantable iNSCs that can be broadly applied for neurological disorders in a safe, efficient, and patient-specific manner.

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BCOR Internal Tandem Duplication Expression in Neural Stem Cells Promotes Growth, Invasion, and Expression of PRC2 Targets

International Journal of Molecular Sciences ◽

10.3390/ijms22083913 ◽

2021 ◽

Vol 22 (8) ◽

pp. 3913

Author(s):

Satoshi Nakata ◽

Ming Yuan ◽

Jeffrey A. Rubens ◽

Ulf D. Kahlert ◽

Jarek Maciaczyk ◽

...

Keyword(s):

Stem Cells ◽

Neural Stem Cells ◽

Tandem Duplication ◽

Target Genes ◽

Cellular Growth ◽

Central Nervous System Tumor ◽

Internal Tandem Duplication ◽

Invasion And Migration ◽

Human Neural Stem Cells ◽

And Migration

Central nervous system tumor with BCL6-corepressor internal tandem duplication (CNS-BCOR ITD) is a malignant entity characterized by recurrent alterations in exon 15 encoding the essential binding domain for the polycomb repressive complex (PRC). In contrast to deletion or truncating mutations seen in other tumors, BCOR expression is upregulated in CNS-BCOR ITD, and a distinct oncogenic mechanism has been suggested. However, the effects of this change on the biology of neuroepithelial cells is poorly understood. In this study, we introduced either wildtype BCOR or BCOR-ITD into human and murine neural stem cells and analyzed them with quantitative RT-PCR and RNA-sequencing, as well as growth, clonogenicity, and invasion assays. In human cells, BCOR-ITD promoted derepression of PRC2-target genes compared to wildtype BCOR. A similar effect was found in clinical specimens from previous studies. However, no growth advantage was seen in the human neural stem cells expressing BCOR-ITD, and long-term models could not be established. In the murine cells, both wildtype BCOR and BCOR-ITD overexpression affected cellular differentiation and histone methylation, but only BCOR-ITD increased cellular growth, invasion, and migration. BCOR-ITD overexpression drives transcriptional changes, possibly due to altered PRC function, and contributes to the oncogenic transformation of neural precursors.

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