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Optimized derivation and functional characterization of 5-HT neurons from human embryonic stem cells

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Optimized derivation and functional characterization of 5-HT neurons from human embryonic stem cells. / Kumar, Manoj; Kaushalya, Sanjeev K; Gressens, Pierre; Maiti, Sudipta; Mani, Shyamala.

In: STEM CELLS AND DEVELOPMENT, Vol. 18, No. 4, 11.05.2009, p. 615-627.

Research output: Contribution to journalArticle

Harvard

Kumar, M, Kaushalya, SK, Gressens, P, Maiti, S & Mani, S 2009, 'Optimized derivation and functional characterization of 5-HT neurons from human embryonic stem cells', STEM CELLS AND DEVELOPMENT, vol. 18, no. 4, pp. 615-627. https://doi.org/10.1089/scd.2008.0181

APA

Kumar, M., Kaushalya, S. K., Gressens, P., Maiti, S., & Mani, S. (2009). Optimized derivation and functional characterization of 5-HT neurons from human embryonic stem cells. STEM CELLS AND DEVELOPMENT, 18(4), 615-627. https://doi.org/10.1089/scd.2008.0181

Vancouver

Kumar M, Kaushalya SK, Gressens P, Maiti S, Mani S. Optimized derivation and functional characterization of 5-HT neurons from human embryonic stem cells. STEM CELLS AND DEVELOPMENT. 2009 May 11;18(4):615-627. https://doi.org/10.1089/scd.2008.0181

Author

Kumar, Manoj ; Kaushalya, Sanjeev K ; Gressens, Pierre ; Maiti, Sudipta ; Mani, Shyamala. / Optimized derivation and functional characterization of 5-HT neurons from human embryonic stem cells. In: STEM CELLS AND DEVELOPMENT. 2009 ; Vol. 18, No. 4. pp. 615-627.

Bibtex Download

@article{4e1ca7f0c151410087ec072ddd75456e,
title = "Optimized derivation and functional characterization of 5-HT neurons from human embryonic stem cells",
abstract = "The ability to study the characteristics of serotonin release from human serotonergic neurons is valuable both in terms of understanding disease pathology and in trying to understand how drugs that affect the serotonergic system alter neurotransmitter release. There is, however, no good in vitro system to model human serotonergic neurons. Although human embryonic stem (hES) cells offer an attractive model system, the derivation of serotonergic neurons from these cells has remained at a low efficiency. To address this problem, Nestin positive precursors from HUES7 hES cell line were first generated. These Nestin positive cells when terminally differentiated gave rise to 20% MAP-2 positive neurons. A high percentage (>40%) of these neurons could be converted to serotonergic neurons. These serotonergic neurons expressed both serotonin and the neuron-specific tryptophan hydroxylase enzyme. In addition, they expressed several of the transcription factors that have been associated with serotonergic differentiation including Mash1 and Pet1. Finally, during the process of neuronal differentiation, the serotonin content, the localization of serotonin vesicles, and their ability to release serotonin following depolarization was characterized using a live cell serotonin imaging technique based on three-photon microscopy. Thus, for the first time, we demonstrate the feasibility of characterizing the development and function of human serotonergic neurons in vitro.",
keywords = "Biological Markers, Cell Differentiation, Cells, Cultured, Cryopreservation, Embryonic Stem Cells, Exocytosis, Fibroblast Growth Factor 2, Humans, Intermediate Filament Proteins, Microtubule-Associated Proteins, Nerve Tissue Proteins, Nestin, Neurons, Phenotype, Progesterone, Putrescine, Serotonin, Tryptophan Hydroxylase",
author = "Manoj Kumar and Kaushalya, {Sanjeev K} and Pierre Gressens and Sudipta Maiti and Shyamala Mani",
year = "2009",
month = may,
day = "11",
doi = "10.1089/scd.2008.0181",
language = "English",
volume = "18",
pages = "615--627",
journal = "STEM CELLS AND DEVELOPMENT",
issn = "1547-3287",
publisher = "Mary Ann Liebert Inc.",
number = "4",

}

RIS (suitable for import to EndNote) Download

TY - JOUR

T1 - Optimized derivation and functional characterization of 5-HT neurons from human embryonic stem cells

AU - Kumar, Manoj

AU - Kaushalya, Sanjeev K

AU - Gressens, Pierre

AU - Maiti, Sudipta

AU - Mani, Shyamala

PY - 2009/5/11

Y1 - 2009/5/11

N2 - The ability to study the characteristics of serotonin release from human serotonergic neurons is valuable both in terms of understanding disease pathology and in trying to understand how drugs that affect the serotonergic system alter neurotransmitter release. There is, however, no good in vitro system to model human serotonergic neurons. Although human embryonic stem (hES) cells offer an attractive model system, the derivation of serotonergic neurons from these cells has remained at a low efficiency. To address this problem, Nestin positive precursors from HUES7 hES cell line were first generated. These Nestin positive cells when terminally differentiated gave rise to 20% MAP-2 positive neurons. A high percentage (>40%) of these neurons could be converted to serotonergic neurons. These serotonergic neurons expressed both serotonin and the neuron-specific tryptophan hydroxylase enzyme. In addition, they expressed several of the transcription factors that have been associated with serotonergic differentiation including Mash1 and Pet1. Finally, during the process of neuronal differentiation, the serotonin content, the localization of serotonin vesicles, and their ability to release serotonin following depolarization was characterized using a live cell serotonin imaging technique based on three-photon microscopy. Thus, for the first time, we demonstrate the feasibility of characterizing the development and function of human serotonergic neurons in vitro.

AB - The ability to study the characteristics of serotonin release from human serotonergic neurons is valuable both in terms of understanding disease pathology and in trying to understand how drugs that affect the serotonergic system alter neurotransmitter release. There is, however, no good in vitro system to model human serotonergic neurons. Although human embryonic stem (hES) cells offer an attractive model system, the derivation of serotonergic neurons from these cells has remained at a low efficiency. To address this problem, Nestin positive precursors from HUES7 hES cell line were first generated. These Nestin positive cells when terminally differentiated gave rise to 20% MAP-2 positive neurons. A high percentage (>40%) of these neurons could be converted to serotonergic neurons. These serotonergic neurons expressed both serotonin and the neuron-specific tryptophan hydroxylase enzyme. In addition, they expressed several of the transcription factors that have been associated with serotonergic differentiation including Mash1 and Pet1. Finally, during the process of neuronal differentiation, the serotonin content, the localization of serotonin vesicles, and their ability to release serotonin following depolarization was characterized using a live cell serotonin imaging technique based on three-photon microscopy. Thus, for the first time, we demonstrate the feasibility of characterizing the development and function of human serotonergic neurons in vitro.

KW - Biological Markers

KW - Cell Differentiation

KW - Cells, Cultured

KW - Cryopreservation

KW - Embryonic Stem Cells

KW - Exocytosis

KW - Fibroblast Growth Factor 2

KW - Humans

KW - Intermediate Filament Proteins

KW - Microtubule-Associated Proteins

KW - Nerve Tissue Proteins

KW - Nestin

KW - Neurons

KW - Phenotype

KW - Progesterone

KW - Putrescine

KW - Serotonin

KW - Tryptophan Hydroxylase

U2 - 10.1089/scd.2008.0181

DO - 10.1089/scd.2008.0181

M3 - Article

C2 - 18800863

VL - 18

SP - 615

EP - 627

JO - STEM CELLS AND DEVELOPMENT

JF - STEM CELLS AND DEVELOPMENT

SN - 1547-3287

IS - 4

ER -

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