A functional human motor unit platform engineered from human embryonic stem cells and immortalized skeletal myoblasts

Marwah Abd Al Samid,1 Jamie S McPhee,2 Jasdeep Saini,1 Tristan R McKay,1 Lorna M Fitzpatrick,1 Kamel Mamchaoui,3 Anne Bigot,3 Vincent Mouly,3 Gillian Butler-Browne,3 Nasser Al-Shanti1 1Healthcare Science Research Institute, School of Healthcare Science, Manchester Metropolitan University, Manchester...

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Autores principales:	Abd Al Samid M, McPhee JS, Saini J, McKay TR, Fitzpatrick LM, Mamchaoui K, Bigot A, Mouly V, Butler-Browne G, Al-Shanti N
Formato:	article
Lenguaje:	EN
Publicado:	Dove Medical Press 2018
Materias:	motor unit neuromuscular junctions human Embryonic Stem cells Neuronal progenitor cells human myoblasts Cytology QH573-671
Acceso en línea:	https://doaj.org/article/554da88af55441119672c7ea10ea2cf6
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spelling	oai:doaj.org-article:554da88af55441119672c7ea10ea2cf62021-12-02T08:10:57ZA functional human motor unit platform engineered from human embryonic stem cells and immortalized skeletal myoblasts1178-6957https://doaj.org/article/554da88af55441119672c7ea10ea2cf62018-11-01T00:00:00Zhttps://www.dovepress.com/a-functional-human-motor-unit-platform-engineered-from-human-embryonic-peer-reviewed-article-SCCAAhttps://doaj.org/toc/1178-6957Marwah Abd Al Samid,1 Jamie S McPhee,2 Jasdeep Saini,1 Tristan R McKay,1 Lorna M Fitzpatrick,1 Kamel Mamchaoui,3 Anne Bigot,3 Vincent Mouly,3 Gillian Butler-Browne,3 Nasser Al-Shanti1 1Healthcare Science Research Institute, School of Healthcare Science, Manchester Metropolitan University, Manchester, UK; 2Department of Sport and Exercise Science, Manchester Metropolitan University, Manchester, UK; 3Center for Research in Myology, Sorbonne Université-INSERM, Paris, France Background: Although considerable research on neuromuscular junctions (NMJs) has been conducted, the prospect of in vivo NMJ studies is limited and these studies are challenging to implement. Therefore, there is a clear unmet need to develop a feasible, robust, and physiologically relevant in vitro NMJ model. Objective: We aimed to establish a novel functional human NMJs platform, which is serum and neural complex media/neural growth factor-free, using human immortalized myoblasts and human embryonic stem cells (hESCs)-derived neural progenitor cells (NPCs) that can be used to understand the mechanisms of NMJ development and degeneration. Methods: Immortalized human myoblasts were co-cultured with hESCs derived committed NPCs. Over the course of the 7 days myoblasts differentiated into myotubes and NPCs differentiated into motor neurons. Results: Neuronal axon sprouting branched to form multiple NMJ innervation sites along the myotubes and the myotubes showed extensive, spontaneous contractile activity. Choline acetyltransferase and βIII-tubulin immunostaining confirmed that the NPCs had matured into cholinergic motor neurons. Postsynaptic site of NMJs was further characterized by staining dihydropyridine receptors, ryanodine receptors, and acetylcholine receptors by α-bungarotoxin. Conclusion: We established a functional human motor unit platform for in vitro investigations. Thus, this co-culture system can be used as a novel platform for 1) drug discovery in the treatment of neuromuscular disorders, 2) deciphering vital features of NMJ formation, regulation, maintenance, and repair, and 3) exploring neuromuscular diseases, age-associated degeneration of the NMJ, muscle aging, and diabetic neuropathy and myopathy. Keywords: motor unit, neuromuscular junctions, human embryonic stem cells, neuronal progenitor cells, human myoblastsAbd Al Samid MMcPhee JSSaini JMcKay TRFitzpatrick LMMamchaoui KBigot AMouly VButler-Browne GAl-Shanti NDove Medical Pressarticlemotor unitneuromuscular junctionshuman Embryonic Stem cellsNeuronal progenitor cellshuman myoblastsCytologyQH573-671ENStem Cells and Cloning: Advances and Applications, Vol Volume 11, Pp 85-93 (2018)
institution	DOAJ
collection	DOAJ
language	EN
topic	motor unit neuromuscular junctions human Embryonic Stem cells Neuronal progenitor cells human myoblasts Cytology QH573-671
spellingShingle	motor unit neuromuscular junctions human Embryonic Stem cells Neuronal progenitor cells human myoblasts Cytology QH573-671 Abd Al Samid M McPhee JS Saini J McKay TR Fitzpatrick LM Mamchaoui K Bigot A Mouly V Butler-Browne G Al-Shanti N A functional human motor unit platform engineered from human embryonic stem cells and immortalized skeletal myoblasts
description	Marwah Abd Al Samid,1 Jamie S McPhee,2 Jasdeep Saini,1 Tristan R McKay,1 Lorna M Fitzpatrick,1 Kamel Mamchaoui,3 Anne Bigot,3 Vincent Mouly,3 Gillian Butler-Browne,3 Nasser Al-Shanti1 1Healthcare Science Research Institute, School of Healthcare Science, Manchester Metropolitan University, Manchester, UK; 2Department of Sport and Exercise Science, Manchester Metropolitan University, Manchester, UK; 3Center for Research in Myology, Sorbonne Université-INSERM, Paris, France Background: Although considerable research on neuromuscular junctions (NMJs) has been conducted, the prospect of in vivo NMJ studies is limited and these studies are challenging to implement. Therefore, there is a clear unmet need to develop a feasible, robust, and physiologically relevant in vitro NMJ model. Objective: We aimed to establish a novel functional human NMJs platform, which is serum and neural complex media/neural growth factor-free, using human immortalized myoblasts and human embryonic stem cells (hESCs)-derived neural progenitor cells (NPCs) that can be used to understand the mechanisms of NMJ development and degeneration. Methods: Immortalized human myoblasts were co-cultured with hESCs derived committed NPCs. Over the course of the 7 days myoblasts differentiated into myotubes and NPCs differentiated into motor neurons. Results: Neuronal axon sprouting branched to form multiple NMJ innervation sites along the myotubes and the myotubes showed extensive, spontaneous contractile activity. Choline acetyltransferase and βIII-tubulin immunostaining confirmed that the NPCs had matured into cholinergic motor neurons. Postsynaptic site of NMJs was further characterized by staining dihydropyridine receptors, ryanodine receptors, and acetylcholine receptors by α-bungarotoxin. Conclusion: We established a functional human motor unit platform for in vitro investigations. Thus, this co-culture system can be used as a novel platform for 1) drug discovery in the treatment of neuromuscular disorders, 2) deciphering vital features of NMJ formation, regulation, maintenance, and repair, and 3) exploring neuromuscular diseases, age-associated degeneration of the NMJ, muscle aging, and diabetic neuropathy and myopathy. Keywords: motor unit, neuromuscular junctions, human embryonic stem cells, neuronal progenitor cells, human myoblasts
format	article
author	Abd Al Samid M McPhee JS Saini J McKay TR Fitzpatrick LM Mamchaoui K Bigot A Mouly V Butler-Browne G Al-Shanti N
author_facet	Abd Al Samid M McPhee JS Saini J McKay TR Fitzpatrick LM Mamchaoui K Bigot A Mouly V Butler-Browne G Al-Shanti N
author_sort	Abd Al Samid M
title	A functional human motor unit platform engineered from human embryonic stem cells and immortalized skeletal myoblasts
title_short	A functional human motor unit platform engineered from human embryonic stem cells and immortalized skeletal myoblasts
title_full	A functional human motor unit platform engineered from human embryonic stem cells and immortalized skeletal myoblasts
title_fullStr	A functional human motor unit platform engineered from human embryonic stem cells and immortalized skeletal myoblasts
title_full_unstemmed	A functional human motor unit platform engineered from human embryonic stem cells and immortalized skeletal myoblasts
title_sort	functional human motor unit platform engineered from human embryonic stem cells and immortalized skeletal myoblasts
publisher	Dove Medical Press
publishDate	2018
url	https://doaj.org/article/554da88af55441119672c7ea10ea2cf6
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A functional human motor unit platform engineered from human embryonic stem cells and immortalized skeletal myoblasts

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