TY - JOUR TI - A 3D culture model of innervated human skeletal muscle enables studies of the adult neuromuscular junction AU - Afshar Bakooshli, Mohsen AU - Lippmann, Ethan S AU - Mulcahy, Ben AU - Iyer, Nisha AU - Nguyen, Christine T AU - Tung, Kayee AU - Stewart, Bryan A AU - van den Dorpel, Hubrecht AU - Fuehrmann, Tobias AU - Shoichet, Molly AU - Bigot, Anne AU - Pegoraro, Elena AU - Ahn, Henry AU - Ginsberg, Howard AU - Zhen, Mei AU - Ashton, Randolph Scott AU - Gilbert, Penney M A2 - Brack, Andrew A2 - Bronner, Marianne E A2 - Brack, Andrew VL - 8 PY - 2019 DA - 2019/05/14 SP - e44530 C1 - eLife 2019;8:e44530 DO - 10.7554/eLife.44530 UR - https://doi.org/10.7554/eLife.44530 AB - 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. KW - 3D co-culture KW - neuromuscular junction KW - skeletal muscle KW - motor neuron KW - myasthenia gravis KW - acetylcholine receptor subunit epsilon JF - eLife SN - 2050-084X PB - eLife Sciences Publications, Ltd ER -