Table 1.
Established in vitro models of neuromuscular diseases. Focus on myoinflammatory, muscular dystrophy, and neuromuscular disease entities.
| Cell Types | Cell Origin | Method | Targeted Mechanism | Disease Model | Novelty Findings | Ref. | |
|---|---|---|---|---|---|---|---|
| Myoin-flammation | T-cells/skeletal myocytes | Primary cells from PM/IBM patients | Monolayer Co-Culture |
Myoinflammation | Myositis (PM, IBM) | Antigen presentation on muscle cells | [99] |
| CD4+ and CD8+ (null) t-cells/autologous skeletal myocytes | Primary cells from PM patients | Monolayer Co-Culture | Myoinflammation | Polymyositis | CD28(null) cells present key effector cells in Polymyositis |
[100] | |
| H2K bOVA- skeletal myocytes/OT-I CD8 + T cells | OVA-specific class I restricted T cell receptor transgenic mice | Monolayer Co-Culture | Myoinflammation/T-cell cytotoxicity | Polymyositis | Invasion of T-cells into myotubes, death of invaded myotubes prior to non-invaded cells | [101] | |
| Dendritic cells/macrophages/skeletal myocytes | Primary cells from myositis patients | Monolayer Co-Culture | Myoinflammation | Myositis | Modulating effect of myoblasts on antigen presenting cells | [102] | |
| Skeletal myocytes | Primary cells from healthy donors | 3D myobundle | Myoinflammation/IFN-γ–induced myopathy | Myositis | Direct IFN-γ-induced muscle weakness, counteracted by exercise-mimetic and JAK/STAT inhibitors |
[103] | |
| Inherited myopathies | Skeletal myocytes | iPSCs derived from DMD patients and control | Monolayer Monoculture | Muscular dystrophy | Duchenne | Morphological and physiological comparable myotubes were able to be differentiated from DMD and control; electric stimulation caused Ca²+-overflow only in DMD-myotubes, this was attenuated after dystrophin restoration through exon-skipping | [104] |
| Skeletal myocytes | Patient-derived iPSCs and genetic correction | Monolayer Monoculture | Restoration of dystrophin protein | Duchenne | Exon skipping, frameshifting, and exon knock-in; exon knock-in was the most effective approach for dystrophin restoration; iPSC-derived skeletal muscle cells with restored protein expression | [105] | |
| Skeletal myocytes | iPSCs of patients with Infantile onset Pompe Disease (IOPD)/healthy controls | Monolayer Monoculture | Lysosomal glycogen accumulation through defect of lysosomal acid α-glucosidase (GAA) | Infantile onset Pompe Disease (IOPD) | Lysosomal glycogen accumulation was dose-dependently rescued by rhGAA; mTOR1-activity is impaired in IOPD with disturbance of energy homeostasis and suppressed mitochondrial oxidative function | [106] | |
| Skeletal myocytes | Human Pompe Disease (PD) iPSCs | Monolayer Monoculture | Lysosomal glycogen accumulation through defect of GAA | Pompe Disease (PD) | Abnormal lysosomal biogenesis is associated with muscular pathology of PD, EB gene transfer is effective as an add-on strategy to GAA gene transfer | [107] | |
| Skeletal myocytes | iPSCs from DMD patients and corrected isogenic iPSCs | Monolayer Monoculture | Muscular dystrophy | Duchenne | Establishment of a human “DMD-in-a-dish” model using DMD-hiPSC-derived myoblasts; disease-related phenotyping with patient-to-patient variability including aberrant expression of inflammation or immune-response genes and collagens, increased BMP/TGFβ signaling, and reduced fusion competence; genetic correction and pharmacological “dual-SMAD” inhibition rescued the genetically corrected isogenic myoblasts forming multi-nucleated myotubes | [85] | |
| Skeletal myofibers | Isogenic DMD mutant cell lines | Monolayer Monoculture | Muscular dystrophy | Duchenne | Improved myofiber maturation from human pluripotent cells in vitro; recapitulation of classical DMD phenotypes in isogenic DMD-mutant iPSC lines; rescue of contractile force, fusion, and branching defects by prednisolone | [108] | |
| Skeletal myocytes | DMD patient-derived iPSC | Monolayer Monoculture | Muscular dystrophy | Duchenne | Generation of contractile human skeletal muscle cells from DMD patient-derived hiPSC based on the inducible expression of MyoD and BAF60C; DMD iPSC-derived myotubes exhibit constitutive activation of TGFβ-SMAD2/3 signaling as well as the deregulated response to pathogenic stimuli, e.g., ECM-derived signals or mechanical cues | [109] | |
| Skeletal myocytes | DMD patient-derived ESC and iPSC, Primary cells from healthy and DMD patients | Monolayer Monoculture | Muscular dystrophy | Duchenne | Transcriptomic evidence of DMD onset before entry into the skeletal muscle compartment during iPSC differentiation; dysregulation of mitochondrial genes identified as one of the earliest detectable changes; early induction of Sonic hedgehog (SSH) signaling pathway, followed by collagens as well as fibrosis-related genes, suggesting the existence of an intrinsic fibrotic process driven by DMD muscle cells. | [110] | |
| Skeletal myocytes/ECs/PCs/SMI32+neurons | hPSCs of healthy donors, Duchenne, LGMD2D and LMNA-related dystrophies | 3D Co-Culture | Muscular dystrophy | Duchenne, LGMD2D and LMNA-related dystrophies | Stable 3D muscle construct of four isogenic cell types, derived from identical hPSCs; detection of muscle-specific as well as disease-related features, | [91] | |
| Skeletal myocytes | Primary cells from healthy and DMD patients | Functionalized monolayer | Muscular dystrophy | Duchenne | Studying of muscle formation and function in functionalized monolayer platform using myoblasts from healthy and DMD patients; impaired polarization with respect to the underlying ECM observed in DMD myoblasts; reduced contractile force | [111] | |
| Neuro-muscular Junction | C2C12 myoblasts/PC12 cells | - | Monolayer Co-culture | Neuron-muscle interaction | - | PC12 cells possess a synergistic effect on C2C12 differentiation | [112] |
| Myofibers/motoneuron | iPSCs | 3D PDMS scaffold | Synaptogenesis | Myasthenia gravis (MG) | Functional connection between motoneuron endplates and myofibers was proven; in the 3D setting accelerated innervation, increased myofiber maturation compared to 2D; MG phenotype was inducible | [113] | |
| Motoneuron-spheroids/myofiber bundles | NSCs/hESCs/iPSC iPSC from a patient with sporadic ALS |
Organ-on-a-chip-model | Synaptogenesis, Drug testing | ALS | Formation of functional NMJ; ALS phenotype with reduced muscle contraction force, neurite regression, and muscle atrophy was contrivable; model feasible for drug testing approaches |
[114] | |
| Organoids resembling the cerebral cortex or the hindbrain/spinal cord/human muscle spheroids | iPSC and primary skeletal myoblasts | 3D cortico-motor-assembloid | Formation of the cortico-motor circuit | - | Cortical controlled muscle contraction was detectable in hPSC derived specific spheroids through relevant neuromuscular connections upon self-assembly; assembloids were stable over several weeks | [115] | |
| Spinal cord neu-rons/skeletal myocytes | hPSC | Neuromuscu-lar Organoids (NMO) | Simultaneous development of spinal cord and muscle compartment in complex 3D organoids | MG | First neuro-muscular organoid model-system that proved highly repro-ducible be-tween exper-iments and different PSC-lines and showed con-tractile activi-ty through functional neuromuscu-lar junctions; MG pheno-type was inducible through ex-posure to autoantibod-ies from MG-patients | [116] | |
| iPSC-derived Motoneurons/skeletal myocytes | iPSC and primary skel-etal my-oblasts | 2D cham-bered co-culture sys-tem | Neuron-muscle inter-action | ALS | Integration of motoneurons derived from ALS-patients’ iPSCs and human skele-tal muscle in chambered co-culture system to develop a functional NMJ model providing a platform to study ALS and being adaptable to patient-specific mod-els | [117] | |
| iPSC-derived Motoneurons/skeletal myocytes | iPSC and primary skel-etal my-oblasts | Chambered co-culture system | Simulation of MG disease mechanisms, drug devel-opment | MG | Functional in vitro MG-model mim-icking reduc-tion in func-tional nA-ChRs at NMJ, decreased NMJ stability, complement activation and blocking of neuromus-cular trans-mission, fea-sible for drug testing | [118] |