De novo revertant fiber formation and therapy testing in a 3D culture model of Duchenne muscular dystrophy skeletal muscle.

摘要: The biological basis of Duchenne muscular dystrophy (DMD) pathology is only partially characterized and there are still few disease-modifying therapies available, therein underlying the value strategies to model study DMD. Dystrophin, causative gene DMD, responsible for linking cytoskeleton muscle fibers extracellular matrix beyond sarcolemma. We posited that disease-associated phenotypes not yet captured by two-dimensional culture methods would arise generating multinucleated cells within a three-dimensional (3D) environment. Herein we report produce 3D human skeletal microtissues (hMMTs) using clonal, immortalized myoblast lines established from healthy DMD donors. also protocols evaluate hMMT self-organization myotube maturation, as well calcium handling, force generation, membrane stability (i.e., creatine kinase activity Evans blue dye permeability) contractile apparatus organization following electrical-stimulation. In examining hMMTs generated with cell line wherein dystrophin possessed duplication exon 2, observed rare dystrophin-positive myotubes, which were seen in 2D cultures. Further, show treating β1-integrin activating antibody, improves maturation stability. Hence, myoblast-derived offer pre-clinical system investigate therapeutic duplicated 2 skipping or those protect contraction-induced injury. STATEMENT OF SIGNIFICANCE: : progressive muscle-wasting disorder caused mutation gene. no cure this fatal disease. Here method myoblasts. Morphological functional assessment revealed DMD-associated pathophysiology including impaired handling presence revertant cells, feature many patients has been recapitulated prior report. further demonstrate "DMD dish" can be used assay test putative mechanism action.