دورية أكاديمية
Generation of Human Motor Units with Functional Neuromuscular Junctions in Microfluidic Devices.
| العنوان: | Generation of Human Motor Units with Functional Neuromuscular Junctions in Microfluidic Devices. |
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| المؤلفون: | Stoklund Dittlau K; Department of Neurosciences, Experimental Neurology, and Leuven Brain Institute, KU Leuven - University of Leuven; VIB Center for Brain & Disease Research, Laboratory of Neurobiology., Krasnow EN; Department of Neurosciences, Experimental Neurology, and Leuven Brain Institute, KU Leuven - University of Leuven; VIB Center for Brain & Disease Research, Laboratory of Neurobiology., Fumagalli L; Department of Neurosciences, Experimental Neurology, and Leuven Brain Institute, KU Leuven - University of Leuven; VIB Center for Brain & Disease Research, Laboratory of Neurobiology., Vandoorne T; Department of Neurosciences, Experimental Neurology, and Leuven Brain Institute, KU Leuven - University of Leuven; VIB Center for Brain & Disease Research, Laboratory of Neurobiology., Baatsen P; VIB Center for Brain & Disease Research, Research Group Molecular Neurobiology, ; VIB Bio Imaging Core, KU Leuven - University of Leuven., Kerstens A; VIB Center for Brain & Disease Research, Research Group Molecular Neurobiology, ; VIB Bio Imaging Core, KU Leuven - University of Leuven., Giacomazzi G; Department of Development and Regeneration, Stem Cell and Developmental Biology, KU Leuven - University of Leuven., Pavie B; VIB Center for Brain & Disease Research, Research Group Molecular Neurobiology, ; VIB Bio Imaging Core, KU Leuven - University of Leuven., Rossaert E; Department of Neurosciences, Experimental Neurology, and Leuven Brain Institute, KU Leuven - University of Leuven; VIB Center for Brain & Disease Research, Laboratory of Neurobiology., Beckers J; Department of Neurosciences, Experimental Neurology, and Leuven Brain Institute, KU Leuven - University of Leuven; VIB Center for Brain & Disease Research, Laboratory of Neurobiology., Sampaolesi M; Department of Development and Regeneration, Stem Cell and Developmental Biology, KU Leuven - University of Leuven., Van Damme P; Department of Neurosciences, Experimental Neurology, and Leuven Brain Institute, KU Leuven - University of Leuven; VIB Center for Brain & Disease Research, Laboratory of Neurobiology, ; Department of Neurology, University Hospitals Leuven., Van Den Bosch L; Department of Neurosciences, Experimental Neurology, and Leuven Brain Institute, KU Leuven - University of Leuven; VIB Center for Brain & Disease Research, Laboratory of Neurobiology, ; ludo.vandenbosch@kuleuven.be. |
| المصدر: | Journal of visualized experiments : JoVE [J Vis Exp] 2021 Sep 07 (175). Date of Electronic Publication: 2021 Sep 07. |
| نوع المنشور: | Journal Article; Research Support, Non-U.S. Gov't; Video-Audio Media |
| اللغة: | English |
| بيانات الدورية: | Publisher: MYJoVE Corporation Country of Publication: United States NLM ID: 101313252 Publication Model: Electronic Cited Medium: Internet ISSN: 1940-087X (Electronic) Linking ISSN: 1940087X NLM ISO Abbreviation: J Vis Exp Subsets: MEDLINE |
| أسماء مطبوعة: | Original Publication: [Boston, Mass. : MYJoVE Corporation, 2006]- |
| مواضيع طبية MeSH: | Induced Pluripotent Stem Cells* , Lab-On-A-Chip Devices*, Humans ; Motor Neurons ; Muscle, Skeletal ; Neuromuscular Junction |
| مستخلص: | Neuromuscular junctions (NMJs) are specialized synapses between the axon of the lower motor neuron and the muscle facilitating the engagement of muscle contraction. In motor neuron disorders, such as amyotrophic lateral sclerosis (ALS) and spinal muscular atrophy (SMA), NMJs degenerate, resulting in muscle atrophy and progressive paralysis. The underlying mechanism of NMJ degeneration is unknown, largely due to the lack of translatable research models. This study aimed to create a versatile and reproducible in vitro model of a human motor unit with functional NMJs. Therefore, human induced pluripotent stem cell (hiPSC)-derived motor neurons and human primary mesoangioblast (MAB)-derived myotubes were co-cultured in commercially available microfluidic devices. The use of fluidically isolated micro-compartments allows for the maintenance of cell-specific microenvironments while permitting cell-to-cell contact through microgrooves. By applying a chemotactic and volumetric gradient, the growth of motor neuron-neurites through the microgrooves promoting myotube interaction and the formation of NMJs were stimulated. These NMJs were identified immunocytochemically through co-localization of motor neuron presynaptic marker synaptophysin (SYP) and postsynaptic acetylcholine receptor (AChR) marker α-bungarotoxin (Btx) on myotubes and characterized morphologically using scanning electron microscopy (SEM). The functionality of the NMJs was confirmed by measuring calcium responses in myotubes upon depolarization of the motor neurons. The motor unit generated using standard microfluidic devices and stem cell technology can aid future research focusing on NMJs in health and disease. |
| تواريخ الأحداث: | Date Created: 20210927 Date Completed: 20211015 Latest Revision: 20211015 |
| رمز التحديث: | 20240628 |
| DOI: | 10.3791/62959 |
| PMID: | 34570099 |
| قاعدة البيانات: | MEDLINE |
| تدمد: | 1940-087X |
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| DOI: | 10.3791/62959 |