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...
| Main Authors: | , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Dove Medical Press
2018-11-01
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| Series: | Stem Cells and Cloning: Advances and Applications |
| Subjects: | |
| Online Access: | https://www.dovepress.com/a-functional-human-motor-unit-platform-engineered-from-human-embryonic-peer-reviewed-article-SCCAA |
| _version_ | 1818739233219674112 |
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| 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 |
| collection | DOAJ |
| 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 |
| first_indexed | 2024-12-18T01:21:34Z |
| format | Article |
| id | doaj.art-554da88af55441119672c7ea10ea2cf6 |
| institution | Directory Open Access Journal |
| issn | 1178-6957 |
| language | English |
| last_indexed | 2024-12-18T01:21:34Z |
| publishDate | 2018-11-01 |
| publisher | Dove Medical Press |
| record_format | Article |
| series | Stem Cells and Cloning: Advances and Applications |
| spelling | doaj.art-554da88af55441119672c7ea10ea2cf62022-12-21T21:25:50ZengDove Medical PressStem Cells and Cloning: Advances and Applications1178-69572018-11-01Volume 11859342186A functional human motor unit platform engineered from human embryonic stem cells and immortalized skeletal myoblastsAbd Al Samid MMcPhee JSSaini JMcKay TRFitzpatrick LMMamchaoui KBigot AMouly VButler-Browne GAl-Shanti NMarwah 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 myoblastshttps://www.dovepress.com/a-functional-human-motor-unit-platform-engineered-from-human-embryonic-peer-reviewed-article-SCCAAmotor unitneuromuscular junctionshuman Embryonic Stem cellsNeuronal progenitor cellshuman myoblasts |
| spellingShingle | 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 Stem Cells and Cloning: Advances and Applications motor unit neuromuscular junctions human Embryonic Stem cells Neuronal progenitor cells human myoblasts |
| title | 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_short | 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 |
| topic | motor unit neuromuscular junctions human Embryonic Stem cells Neuronal progenitor cells human myoblasts |
| url | https://www.dovepress.com/a-functional-human-motor-unit-platform-engineered-from-human-embryonic-peer-reviewed-article-SCCAA |
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