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Differentiation of Human Embryonic Stem Cells to Sympathetic Neurons: A Potential Model for Understanding Neuroblastoma Pathogenesis

Carr-Wilkinson, Jane, Prathalingam, Nilendran, Deepali, Pal, Moad, Mohammad, Lee, Natalie, Sundaresh, Aishwarya, Forgham, Helen, James, Peter, Herbert, Mary, Lako, Majlinda and Tweddle, Deborah A (2018) Differentiation of Human Embryonic Stem Cells to Sympathetic Neurons: A Potential Model for Understanding Neuroblastoma Pathogenesis. Stem Cells International. ISSN 0378-4274

Item Type: Article

Abstract

Background and Aims: Previous studies modelling human neural crest differentiation from stem cells have resulted in a low yield of sympathetic neurons. Our aim was to optimise a method for the differentiation of human embryonic stem cells (hESCs) to sympathetic neuron-like cells (SN) to model normal human SNS development.

Results: Using stromal-derived inducing activity (SDIA) of PA6 cells plus BMP4 and B27 supplements, the H9 hESC line was differentiated to neural crest stem-like cells and SN-like cells. After 7 days of PA6 cell coculture, mRNA expression of SNAIL and SOX-9 neural crest specifier genes and the neural marker peripherin (PRPH) increased. Expression of the pluripotency marker OCT 4 decreased, whereas TP53 and LIN28B expression remained high at levels similar to SHSY5Y and IMR32 neuroblastoma cell lines. A 5-fold increase in the expression of the catecholaminergic marker tyrosine hydroxylase (TH) and the noradrenergic marker dopamine betahydroxylase (DBH) was observed by day 7 of differentiation. Fluorescence-activated cell sorting for the neural crest marker p75, enriched for cells expressing p75, DBH, TH, and PRPH, was more specific than p75 neural crest stem cell (NCSC) microbeads. On day 28 post p75 sorting, dual immunofluorescence identified sympathetic neurons by PRPH and TH copositivity cells in 20% of the cell population. Noradrenergic sympathetic neurons, identified by copositivity for both PHOX2B and DBH, were present in 9.4% ± 5.5% of cells.

Conclusions: We have optimised a method for noradrenergic SNS development using the H9 hESC line to improve our understanding of normal human SNS development and, in a future work, the pathogenesis of neuroblastoma.

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Depositing User: Jane Carr-Wilkinson

Identifiers

Item ID: 10463
Identification Number: https://doi.org/10.1155/2018/4391641
ISSN: 0378-4274
URI: http://sure.sunderland.ac.uk/id/eprint/10463
Official URL: https://www.hindawi.com/journals/sci/2018/4391641/

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Catalogue record

Date Deposited: 22 Mar 2019 13:36
Last Modified: 18 Dec 2019 16:07

Contributors

Author: Jane Carr-Wilkinson
Author: Nilendran Prathalingam
Author: Pal Deepali
Author: Mohammad Moad
Author: Natalie Lee
Author: Aishwarya Sundaresh
Author: Helen Forgham
Author: Peter James
Author: Mary Herbert
Author: Majlinda Lako
Author: Deborah A Tweddle

University Divisions

Faculty of Health Sciences and Wellbeing
Faculty of Health Sciences and Wellbeing > School of Nursing and Health Sciences

Subjects

Sciences > Biomedical Sciences
Sciences > Health Sciences

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