3D human induced pluripotent stem cell–derived bioengineered skeletal muscles for tissue, disease and therapy modeling

Luca Pinton, Moustafa Khedr, Valentina M. Lionello, Shilpita Sarcar, Sara M. Maffioletti, Sumitava Dastidar, Elisa Negroni, SungWoo Choi, Noreen Khokhar, Anne Bigot, John R. Counsell, Andreia Sofia Bernardo, Peter S. Zammit, Francesco Saverio Tedesco

Published: 2023-02-14 DOI: 10.1038/s41596-022-00790-8

induced pluripotent stem cells

bioengineered skeletal muscles

3D hydrogels

muscle pathology

therapy modeling

AI 解读

Supplementary information

Supplementary Information

Supplementary Table 1 and Protocol 1

Supplementary Video 1

Real-time video of DMD hiPSC-derived 3D skeletal muscle construct contractions in response to 10-V electrical stimulation at 1 Hz

Supplementary Video 2

Calcium transients in DMD hiPSC-derived 3D artificial muscles incubated with the fluorescence Ca 2+ indicator Fluo-4AM upon electrical stimulation. During the first 10 s, no electrical stimuli were applied to the muscle. At second 10, the 3D muscle was stimulated with 10 V at 0.33 Hz, and at second 20, the 3D muscle was stimulated with 20 V at 0.33 Hz

Supplementary Video 3

Maximum-intensity projection time-lapse video of 5-chloromethylfluorescein diacetate (CMFDA)-stained HIDEMs (green) deposited on hiPSC-derived 3D muscles (red). 100-µm stacks of muscles were imaged every 12 min for 4 h by using a spinning disk microscope. Scale bar, 100 µm

Supplementary Video 4

3D reconstruction of a 3D artificial muscle imaged with a light-sheet microscope immunostained as detailed in the advanced imaging section of Fig. 2

Supplementary Video 5

3D reconstruction nuclear morphology (lamin A/C in red) and myotubes (eMyHC in green) of a healthy control hiPSC-derived 3D muscle construct shown in Fig. 3. The 3D reconstruction was done by using Imaris software and confocal z-stacks of immunostained artificial muscles. Video reproduced from ref. 29 under Creative Commons license CC BY 4.0

Supplementary Video 6

3D reconstruction nuclear morphology (lamin A/C in red) and myotubes (eMyHC in green) of an LMNA-mutant (R249W) hiPSC-derived 3D muscle construct shown in Fig. 3. The 3D reconstruction was done by using Imaris software and confocal z-stacks of immunostained artificial muscles. Video reproduced from ref. 29 under Creative Commons license CC BY 4.0

References

Supplementary information

Tedesco, F. S., Dellavalle, A., Diaz-Manera, J., Messina, G. & Cossu, G. Repairing skeletal muscle: regenerative potential of skeletal muscle stem cells. J. Clin. Invest. 120, 11–19 (2010).
Wosczyna, M. N. & Rando, T. A. A muscle stem cell support group: coordinated cellular responses in muscle regeneration. Dev. Cell. 46, 135–143 (2018).
Tedesco, F. S., Moyle, L. A. & Perdiguero, E. Muscle interstitial cells: a brief field guide to non-satellite cell populations in skeletal muscle. Methods Mol. Biol. 1556, 129–147 (2017).
Mercuri, E., Bonnemann, C. G. & Muntoni, F. Muscular dystrophies. Lancet 394, 2025–2038 (2019).
Jalal, S., Dastidar, S. & Tedesco, F. S. Advanced models of human skeletal muscle differentiation, development and disease: Three-dimensional cultures, organoids and beyond. Curr. Opin. Cell Biol. 73, 92–104 (2021).
Fatehullah, A., Tan, S. H. & Barker, N. Organoids as an in vitro model of human development and disease. Nat. Cell Biol. 18, 246–254 (2016).
Lancaster, M. A. & Knoblich, J. A. Organogenesis in a dish: modeling development and disease using organoid technologies. Science 345, 1247125 (2014).
Wang, L., Wu, Y., Guo, B. & Ma, P. X. Nanofiber yarn/hydrogel core-shell scaffolds mimicking native skeletal muscle tissue for guiding 3D myoblast alignment, elongation, and differentiation. ACS Nano 9, 9167–9179 (2015).
Shimizu, K. et al. Microfluidic devices for construction of contractile skeletal muscle microtissues. J. Biosci. Bioeng. 119, 212–216 (2015).
Sakar, M. S. et al. Formation and optogenetic control of engineered 3D skeletal muscle bioactuators. Lab Chip 12, 4976–4985 (2012).
Pruller, J., Mannhardt, I., Eschenhagen, T., Zammit, P. S. & Figeac, N. Satellite cells delivered in their niche efficiently generate functional myotubes in three-dimensional cell culture. PLoS One 13, e0202574 (2018).
Morimoto, Y., Kato-Negishi, M., Onoe, H. & Takeuchi, S. Three-dimensional neuron-muscle constructs with neuromuscular junctions. Biomaterials 34, 9413–9419 (2013).
Juhas, M., Engelmayr, G. C. Jr., Fontanella, A. N., Palmer, G. M. & Bursac, N. Biomimetic engineered muscle with capacity for vascular integration and functional maturation in vivo. Proc. Natl. Acad. Sci. USA 111, 5508–5513 (2014).
Heher, P. et al. A novel bioreactor for the generation of highly aligned 3D skeletal muscle-like constructs through orientation of fibrin via application of static strain. Acta Biomater. 24, 251–265 (2015).
Cvetkovic, C. et al. Three-dimensionally printed biological machines powered by skeletal muscle. Proc. Natl. Acad. Sci. USA 111, 10125–10130 (2014).
Corona, B. T., Ward, C. L., Baker, H. B., Walters, T. J. & Christ, G. J. Implantation of in vitro tissue engineered muscle repair constructs and bladder acellular matrices partially restore in vivo skeletal muscle function in a rat model of volumetric muscle loss injury. Tissue Eng. Part A 20, 705–715 (2014).
Chen, H. et al. Enhanced growth and differentiation of myoblast cells grown on E-jet 3D printed platforms. Int. J. Nanomedicine 14, 937–950 (2019).
Capel, A. J. et al. Scalable 3D printed molds for human tissue engineered skeletal muscle. Front. Bioeng. Biotechnol. 7, 20 (2019).
Fusto, A., Moyle, L. A., Gilbert, P. M. & Pegoraro, E. Cored in the act: the use of models to understand core myopathies. Dis. Model. Mech. 12, dmm.041368 (2019).
Trevisan, C. et al. Generation of a functioning and self-renewing diaphragmatic muscle construct. Stem Cells Transl. Med. 8, 858–869 (2019).
Tchao, J. et al. Engineered human muscle tissue from skeletal muscle derived stem cells and induced pluripotent stem cell derived cardiac cells. Int. J. Tissue Eng. 2013, 198762 (2013).
Quarta, M. et al. Bioengineered constructs combined with exercise enhance stem cell-mediated treatment of volumetric muscle loss. Nat. Commun. 8, 15613 (2017).
Powell, C. A., Smiley, B. L., Mills, J. & Vandenburgh, H. H. Mechanical stimulation improves tissue-engineered human skeletal muscle. Am. J. Physiol. Cell Physiol. 283, C1557–C1565 (2002).
Passipieri, J. A. et al. Keratin hydrogel enhances in vivo skeletal muscle function in a rat model of volumetric muscle loss. Tissue Eng. Part A 23, 556–571 (2017).
Madden, L., Juhas, M., Kraus, W. E., Truskey, G. A. & Bursac, N. Bioengineered human myobundles mimic clinical responses of skeletal muscle to drugs. Elife 4, e04885 (2015).
Fuoco, C. et al. In vivo generation of a mature and functional artificial skeletal muscle. EMBO Mol. Med. 7, 411–422 (2015).
Chiron, S. et al. Complex interactions between human myoblasts and the surrounding 3D fibrin-based matrix. PLoS One 7, e36173 (2012).
Bersini, S. et al. Engineering an environment for the study of fibrosis: a 3D human muscle model with endothelium specificity and endomysium. Cell Rep. 25, 3858–3868.e4 (2018).
Afshar Bakooshli, M. et al. A 3D culture model of innervated human skeletal muscle enables studies of the adult neuromuscular junction. Elife 8, e44530 (2019).
Steele-Stallard, H. B. et al. Modeling skeletal muscle laminopathies using human induced pluripotent stem cells carrying pathogenic LMNA mutations. Front. Physiol. 9, 1332 (2018).
Maffioletti, S. M. et al. Three-dimensional human iPSC-derived artificial skeletal muscles model muscular dystrophies and enable multilineage tissue engineering. Cell Rep. 23, 899–908 (2018).
Rao, L., Qian, Y., Khodabukus, A., Ribar, T. & Bursac, N. Engineering human pluripotent stem cells into a functional skeletal muscle tissue. Nat. Commun. 9, 126 (2018).
Osaki, T., Uzel, S. G. M. & Kamm, R. D. Microphysiological 3D model of amyotrophic lateral sclerosis (ALS) from human iPS-derived muscle cells and optogenetic motor neurons. Sci. Adv. 4, eaat5847 (2018).
Faustino Martins, J. M. et al. Self-organizing 3D human trunk neuromuscular organoids. Cell Stem Cell 26, 172–186.e6 (2020).
Relaix, F. & Zammit, P. S. Satellite cells are essential for skeletal muscle regeneration: the cell on the edge returns centre stage. Development 139, 2845–2856 (2012).
Worman, H. J. Nuclear lamins and laminopathies. J. Pathol. 226, 316–325 (2012).
Maffioletti, S. M. et al. Efficient derivation and inducible differentiation of expandable skeletal myogenic cells from human ES and patient-specific iPS cells. Nat. Protoc. 10, 941–958 (2015).
Caron, L. et al. A human pluripotent stem cell model of facioscapulohumeral muscular dystrophy-affected skeletal muscles. Stem Cells Transl. Med. 5, 1145–1161 (2016).
Orlova, V. V. et al. Generation, expansion and functional analysis of endothelial cells and pericytes derived from human pluripotent stem cells. Nat. Protoc. 9, 1514–1531 (2014).
Stacpoole, S. R. et al. Efficient derivation of NPCs, spinal motor neurons and midbrain dopaminergic neurons from hESCs at 3% oxygen. Nat. Protoc. 6, 1229–1240 (2011).
Hall, C. E. et al. Progressive motor neuron pathology and the role of astrocytes in a human stem cell model of VCP-related ALS. Cell Rep. 19, 1739–1749 (2017).
Machado, C. B. et al. In vitro modelling of nerve-muscle connectivity in a compartmentalised tissue culture device. Adv. Biosyst. 3, 1800307 (2019).
Paredes-Redondo, A. et al. Optogenetic modeling of human neuromuscular circuits in Duchenne muscular dystrophy with CRISPR and pharmacological corrections. Sci. Adv. 7, eabi8787 (2021).
Martin, N. R. et al. Neuromuscular junction formation in tissue-engineered skeletal muscle augments contractile function and improves cytoskeletal organization. Tissue Eng. Part A 21, 2595–2604 (2015).
Mills, R. J. et al. Development of a human skeletal micro muscle platform with pacing capabilities. Biomaterials 198, 217–227 (2019).
Afshar, M. E. et al. A 96-well culture platform enables longitudinal analyses of engineered human skeletal muscle microtissue strength. Sci. Rep. 10, 6918 (2020).
Grebenyuk, S. & Ranga, A. Engineering organoid vascularization. Front. Bioeng. Biotechnol. 7, 39 (2019).
de la Puente, P. & Ludena, D. Cell culture in autologous fibrin scaffolds for applications in tissue engineering. Exp. Cell Res. 322, 1–11 (2014).
Ben Yaou, R. et al. International retrospective natural history study of LMNA-related congenital muscular dystrophy. Brain Commun. 3, fcab075 (2021).
Rose, N., et al. Bioengineering a miniaturized in vitro 3D myotube contraction monitoring chip for modelization of muscular dystrophies. Biomater. 293, 121935 (2023).
Tedesco, F. S. et al. Stem cell–mediated transfer of a human artificial chromosome ameliorates muscular dystrophy. Sci. Transl. Med. 3, 96ra78 (2011).
Mendell, J. R. et al. Single-dose gene-replacement therapy for spinal muscular atrophy. New England J. Med. 377, 1713–1722 (2017).
Choi, S., Ferrari, G. & Tedesco, F. S. Cellular dynamics of myogenic cell migration: molecular mechanisms and implications for skeletal muscle cell therapies. EMBO Mol. Med. 12, e12357 (2020).
Choi, S. et al. Assessing and enhancing migration of human myogenic progenitors using directed iPS cell differentiation and advanced tissue modelling. EMBO Mol. Med. 14, e14526 (2022).
Hagemann, C. et al. Soft-lithography on SLA 3D printed moulds for fast, versatile, and accessible high-resolution fabrication of customised multiscale cell culture devices with complex designs Preprint at bioRxiv http://doi.org/10.1101/2022.02.22.481424 (2022).
Loperfido, M., Steele-Stallard, H. B., Tedesco, F. S. & VandenDriessche, T. Pluripotent stem cells for gene therapy of degenerative muscle diseases. Curr. Gene Ther. 15, 364–380 (2015).
Selvaraj, S., Kyba, M. & Perlingeiro, R. C. R. Pluripotent stem cell-based therapeutics for muscular dystrophies. Trends Mol. Med. 25, 803–816 (2019).
Darabi, R. et al. Human ES- and iPS-derived myogenic progenitors restore DYSTROPHIN and improve contractility upon transplantation in dystrophic mice. Cell Stem Cell 10, 610–619 (2012).
Tedesco, F. S. et al. Transplantation of genetically corrected human iPSC-derived progenitors in mice with limb-girdle muscular dystrophy. Sci. Transl. Med. 4, 140ra189 (2012).
Goudenege, S. et al. Myoblasts derived from normal hESCs and dystrophic hiPSCs efficiently fuse with existing muscle fibers following transplantation. Mol. Ther. 20, 2153–2167 (2012).
Shoji, E. et al. Early pathogenesis of Duchenne muscular dystrophy modelled in patient-derived human induced pluripotent stem cells. Sci. Rep. 5, 12831 (2015).
Kim, H. et al. Genomic safe harbor expression of PAX7 for the generation of engraftable myogenic progenitors. Stem Cell Rep. 16, 10–19 (2021).
Albini, S. et al. Epigenetic reprogramming of human embryonic stem cells into skeletal muscle cells and generation of contractile myospheres. Cell Rep. 3, 661–670 (2013).
Hicks, M. R. et al. ERBB3 and NGFR mark a distinct skeletal muscle progenitor cell in human development and hPSCs. Nat. Cell Biol. 20, 46–57 (2018).
Chal, J. et al. Generation of human muscle fibers and satellite-like cells from human pluripotent stem cells in vitro. Nat. Protoc. 11, 1833–1850 (2016).
Borchin, B., Chen, J. & Barberi, T. Derivation and FACS-mediated purification of PAX3+/PAX7+ skeletal muscle precursors from human pluripotent stem cells. Stem Cell Rep. 1, 620–631 (2013).
Hansen, A. et al. Development of a drug screening platform based on engineered heart tissue. Circ. Res. 107, 35–44 (2010).
Sala, L. et al. MUSCLEMOTION: a versatile open software tool to quantify cardiomyocyte and cardiac muscle contraction in vitro and in vivo. Circ. Res. 122, e5–e16 (2018).