| Title: |
A 3D culture model of innervated human skeletal muscle enables studies of the adult neuromuscular junction |
| Authors: |
Afshar Bakooshli, Mohsen; Lippmann, Ethan S; Mulcahy, Ben; Iyer, Nisha; Nguyen, Christine T; Tung, Kayee; Stewart, Bryan A; van den Dorpel, Hubrecht; Fuehrmann, Tobias; Shoichet, Molly; Bigot, Anne; Pegoraro, Elena; Ahn, Henry; Ginsberg, Howard; Zhen, Mei; Ashton, Randolph Scott; Gilbert, Penney M |
| Contributors: |
Natural Sciences and Engineering Research Council of Canada; Krembil Foundation; Toronto Musculoskeletal Centre; University of Toronto; National Institutes of Health; TD Bank Health Research Fellowship; AFM-Téléthon; Canadian Institutes of Health Research; Burroughs Wellcome Fund; U.S. Environmental Protection Agency; Toronto Western Arthritis Program; Ontario Research Fund; Canada Research Chairs; Canada Foundation for Innovation; Canada First Research Excellence Fund; Ontario Institute for Regenerative Medicine; University of Toronto Faculty of Medicine |
| Source: |
eLife ; volume 8 ; ISSN 2050-084X |
| Publisher Information: |
eLife Sciences Publications, Ltd |
| Publication Year: |
2019 |
| Collection: |
eLife (E-Journal - via CrossRef) |
| Description: |
Two-dimensional (2D) human skeletal muscle fiber cultures are ill-equipped to support the contractile properties of maturing muscle fibers. This limits their application to the study of adult human neuromuscular junction (NMJ) development, a process requiring maturation of muscle fibers in the presence of motor neuron endplates. Here we describe a three-dimensional (3D) co-culture method whereby human muscle progenitors mixed with human pluripotent stem cell-derived motor neurons self-organize to form functional NMJ connections. Functional connectivity between motor neuron endplates and muscle fibers is confirmed with calcium imaging and electrophysiological recordings. Notably, we only observed epsilon acetylcholine receptor subunit protein upregulation and activity in 3D co-cultures. Further, 3D co-culture treatments with myasthenia gravis patient sera shows the ease of studying human disease with the system. Hence, this work offers a simple method to model and evaluate adult human NMJ de novo development or disease in culture. |
| Document Type: |
article in journal/newspaper |
| Language: |
English |
| DOI: |
10.7554/elife.44530 |
| Availability: |
https://doi.org/10.7554/elife.44530; https://cdn.elifesciences.org/articles/44530/elife-44530-v1.pdf; https://cdn.elifesciences.org/articles/44530/elife-44530-v1.xml; https://elifesciences.org/articles/44530 |
| Rights: |
http://creativecommons.org/licenses/by/4.0/ ; http://creativecommons.org/licenses/by/4.0/ ; http://creativecommons.org/licenses/by/4.0/ |
| Accession Number: |
edsbas.97AAA911 |
| Database: |
BASE |