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induce hPSCs into myogenic progenitor cells and myoblasts, research and therapy development due to the interspecies
enabling the biofabrication of human skeletal muscle divergence, while organoids can provide an ideal platform
organoids (hSkMOs), and observed sustainable satellite cells that more closely resembles the physiological environment.
that can be activated for repairing damaged muscle tissue Shahriyari et al. generated functional human skeletal
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during the culture process. Starting from human hiPSC, muscle organoids (SMOs) and engineered skeletal muscle
Grass et al. generated neuromesodermal progenitors from hPSCs. Utilizing patient-derived iPSCs with a
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(NMPs) through WNT and FGF signaling activation, and deletion of exons 48–50, they constructed an engineered
subsequently induced differentiation of a portion of NMPs skeletal muscle model with features of Duchenne muscular
that retain mesodermal identity to skeletal myocytes, dystrophy (DMD), and used this model to demonstrate
enabling the construction of neuromuscular organoids. the therapeutic efficacy of CRISPR/Cas9 technology
Bioengineering techniques have been used in organoid in DMD. Gao et al. used iPSCs from C9orf72-ALS
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construction to increase the efficiency of construction patients to generate neuromuscular organoids which
and to improve the homogeneity of the organoids, thereby showed ALS-associated lesion features. On this basis, they
increasing their potential for application. For example, performed drug testing and demonstrated the efficacy of
Li et al. developed functional mouse skeletal muscle GSK2606414 in improving skeletal muscle contraction
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droplet-engineered organoids from mouse gastrocnemius and decreasing the accumulation of poly (glycine-proline)
muscle tissue using cascade tube microfluidics (CTM). dipeptide repeat protein. Organoids can also be used to
Constructed within a shorter duration, the organoids study the effects of various possible pathogenic factors. For
exhibited enhanced maturation and functionality, as well example, Jiang et al. established hSkMOs derived from
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as potential for scalable and reproducible production. Such hPSC that maintained important skeletal muscle features
an organoid production technique has high feasibility and and then exposed them to a 2 Gy dose of radiation and
substantial potential for both fundamental research and observed defects in organoid amplification, differentiation,
therapeutic applications. In addition, recapitulation of and repair. In addition to this, muscle organoids can also
actual physiological condition, mimicry of higher-order be used for drug screening. Svobodova et al. used hiPSC
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architectural and functional complexity, and construction to establish a disease model of DMD in vitro, providing a
of multi-tissue composite organoids are being explored. Yin high-throughput platform for drug screening. Furthermore,
et al. established neuromusculoskeletal organoids from the construction of patient-specific organoids can also
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hPSCs through a co-culture strategy (static-to-spinning facilitate the development of targeted therapies (Figure 2).
culture strategy), which realized the coordinated
development of three distinct tissue domains within a single 2.2. Tendon organoids
organoid, demonstrating the strong self-organization ability
of 3D cell culture systems in vitro (Table 2). 2.2.1. Physiological structure of tendon
Tendon is mainly a collagen fiber bundle structure
2.1.3. Application of skeletal muscle organoid connecting muscle and bone, which is mainly composed
Traditional models adopted in disease research are of type I collagen, and similar to the way skeletal muscle is
primarily 2D cell cultures and animal models, but 2D organized, collagen molecules form procollagen molecules,
cultures are difficult to replicate cell growth in a 3D five procollagen molecules group together to form
environment in vivo, and animal models exhibit inherent microfibrils, which subsequently aggregate into fibrils, and
limitations and inaccuracy in disease mechanism fibrils are combined into collagen fibers to ultimately give

Table 2. Construction of skeletal muscle organoids
Cell source Inducing factor Matrix material References
iPSC bFGF, CHIR99021, Y27632, HGF/IGF Geltrex 32
hPSC HGF, IGF1, FGF2 Growth factor-reduced Matrigel 33
Immortalized myoblasts Human recombinant insulin Hydrogel 34
hiPSC FGF2, CHIR99021, GDF11 Matrigel 36
Primary skeletal muscle cells IGF1 Matrigel 37
hPSC HGF, FGF2 Collagen/Matrigel hydrogel 39
hPSC FGF2 N/A 38
hiPSC IGF, FGF2, HGF, CHIR99021 Matrigel 40
Abbreviations: bFGF: Basic fibroblast growth factor; FGF: Fibroblast growth factor; GDF11: Growth differentiation factor 11; HGF: Hepatocyte
growth factor; hiPSC: Human-induced pluripotent stem cells; hPSC: Human pluripotent stem cells; IGF: Insulin-like growth factor; iPSC: Induced
pluripotent stem cell.

Volume 1 Issue 3 (2025) 4 doi: 10.36922/OR025320025