Generation of kisspeptin-responsive GnRH neurons from human pluripotent stem cells

Ariel Poliandri; Duncan Miller; Sasha Howard; Muriel Nobles; Gerard Ruiz-Babot; Stephen Harmer; Andrew Tinker; Tristan McKay; Leonardo Guasti; Leo Dunkel

doi:10.1016/j.mce.2017.02.030

Generation of kisspeptin-responsive GnRH neurons from human pluripotent stem cells

Ariel Poliandri
, Duncan Miller
, Sasha Howard
, Muriel Nobles
, Gerard Ruiz-Babot
, Stephen Harmer
, Andrew Tinker
, Tristan McKay
, Leonardo Guasti
, Leo Dunkel

Research output: Contribution to journal › Article (Academic Journal) › peer-review

14 Citations (Scopus)

312 Downloads (Pure)

Abstract

GnRH neurons are fundamental for reproduction in all vertebrates, integrating all reproductive inputs. The inaccessibility of human GnRH-neurons has been a major impediment to studying the central control of reproduction and its disorders. Here, we report the efficient generation of kisspeptin responsive GnRH-secreting neurons by directed differentiation of human Embryonic Stem Cells and induced-Pluripotent Stem Cells derived from a Kallman Syndrome patient and a healthy family member. The protocol involves the generation of intermediate Neural Progenitor Cells (NPCs) through long-term Bone morphogenetic protein 4 inhibition, followed by terminal specification of these NPCs in media containing Fibroblast Growth Factor 8 and a NOTCH inhibitor. The resulting GnRH-expressing and -secreting neurons display a neuroendocrine gene expression pattern and present spontaneous calcium transients that can be stimulated by kisspeptin. These in vitro generated GnRH expressing cells provide a new resource for studying the molecular mechanisms underlying the development and function of GnRH neurons.

Original language	English
Pages (from-to)	12-22
Number of pages	11
Journal	Molecular and Cellular Endocrinology
Volume	447
Early online date	21 Feb 2017
DOIs	https://doi.org/10.1016/j.mce.2017.02.030
Publication status	Published - 15 May 2017

Bibliographical note

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

SDG 3 Good Health and Well-being

Keywords

Calcium/metabolism
Cell Differentiation
Cell Line
Gene Expression Profiling
Gene Expression Regulation
Gonadotropin-Releasing Hormone/metabolism
Human Embryonic Stem Cells/metabolism
Humans
Kisspeptins/metabolism
Neural Stem Cells/metabolism
Neurons/metabolism
Pluripotent Stem Cells/metabolism

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10.1016/j.mce.2017.02.030

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Accepted author manuscript, 363 KBLicence: CC BY-NC-ND

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@article{48168c7e18294140ac8ed4eec9be445e,

title = "Generation of kisspeptin-responsive GnRH neurons from human pluripotent stem cells",

abstract = "GnRH neurons are fundamental for reproduction in all vertebrates, integrating all reproductive inputs. The inaccessibility of human GnRH-neurons has been a major impediment to studying the central control of reproduction and its disorders. Here, we report the efficient generation of kisspeptin responsive GnRH-secreting neurons by directed differentiation of human Embryonic Stem Cells and induced-Pluripotent Stem Cells derived from a Kallman Syndrome patient and a healthy family member. The protocol involves the generation of intermediate Neural Progenitor Cells (NPCs) through long-term Bone morphogenetic protein 4 inhibition, followed by terminal specification of these NPCs in media containing Fibroblast Growth Factor 8 and a NOTCH inhibitor. The resulting GnRH-expressing and -secreting neurons display a neuroendocrine gene expression pattern and present spontaneous calcium transients that can be stimulated by kisspeptin. These in vitro generated GnRH expressing cells provide a new resource for studying the molecular mechanisms underlying the development and function of GnRH neurons.",

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author = "Ariel Poliandri and Duncan Miller and Sasha Howard and Muriel Nobles and Gerard Ruiz-Babot and Stephen Harmer and Andrew Tinker and Tristan McKay and Leonardo Guasti and Leo Dunkel",

year = "2017",

month = may,

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doi = "10.1016/j.mce.2017.02.030",

language = "English",

volume = "447",

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journal = "Molecular and Cellular Endocrinology",

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TY - JOUR

T1 - Generation of kisspeptin-responsive GnRH neurons from human pluripotent stem cells

AU - Poliandri, Ariel

AU - Miller, Duncan

AU - Howard, Sasha

AU - Nobles, Muriel

AU - Ruiz-Babot, Gerard

AU - Harmer, Stephen

AU - Tinker, Andrew

AU - McKay, Tristan

AU - Guasti, Leonardo

AU - Dunkel, Leo

PY - 2017/5/15

Y1 - 2017/5/15

N2 - GnRH neurons are fundamental for reproduction in all vertebrates, integrating all reproductive inputs. The inaccessibility of human GnRH-neurons has been a major impediment to studying the central control of reproduction and its disorders. Here, we report the efficient generation of kisspeptin responsive GnRH-secreting neurons by directed differentiation of human Embryonic Stem Cells and induced-Pluripotent Stem Cells derived from a Kallman Syndrome patient and a healthy family member. The protocol involves the generation of intermediate Neural Progenitor Cells (NPCs) through long-term Bone morphogenetic protein 4 inhibition, followed by terminal specification of these NPCs in media containing Fibroblast Growth Factor 8 and a NOTCH inhibitor. The resulting GnRH-expressing and -secreting neurons display a neuroendocrine gene expression pattern and present spontaneous calcium transients that can be stimulated by kisspeptin. These in vitro generated GnRH expressing cells provide a new resource for studying the molecular mechanisms underlying the development and function of GnRH neurons.

AB - GnRH neurons are fundamental for reproduction in all vertebrates, integrating all reproductive inputs. The inaccessibility of human GnRH-neurons has been a major impediment to studying the central control of reproduction and its disorders. Here, we report the efficient generation of kisspeptin responsive GnRH-secreting neurons by directed differentiation of human Embryonic Stem Cells and induced-Pluripotent Stem Cells derived from a Kallman Syndrome patient and a healthy family member. The protocol involves the generation of intermediate Neural Progenitor Cells (NPCs) through long-term Bone morphogenetic protein 4 inhibition, followed by terminal specification of these NPCs in media containing Fibroblast Growth Factor 8 and a NOTCH inhibitor. The resulting GnRH-expressing and -secreting neurons display a neuroendocrine gene expression pattern and present spontaneous calcium transients that can be stimulated by kisspeptin. These in vitro generated GnRH expressing cells provide a new resource for studying the molecular mechanisms underlying the development and function of GnRH neurons.

KW - Calcium/metabolism

KW - Cell Differentiation

KW - Cell Line

KW - Gene Expression Profiling

KW - Gene Expression Regulation

KW - Gonadotropin-Releasing Hormone/metabolism

KW - Human Embryonic Stem Cells/metabolism

KW - Humans

KW - Kisspeptins/metabolism

KW - Neural Stem Cells/metabolism

KW - Neurons/metabolism

KW - Pluripotent Stem Cells/metabolism

U2 - 10.1016/j.mce.2017.02.030

DO - 10.1016/j.mce.2017.02.030

M3 - Article (Academic Journal)

C2 - 28232089

SN - 0303-7207

VL - 447

SP - 12

EP - 22

JO - Molecular and Cellular Endocrinology

JF - Molecular and Cellular Endocrinology

ER -

Generation of kisspeptin-responsive GnRH neurons from human pluripotent stem cells

Abstract

Bibliographical note

UN SDGs

Keywords

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