Advanced models of human skeletal muscle differentiation, development and disease: Three-dimensional cultures, organoids and beyond
Introduction
The skeletal muscle, an architecturally complex tissue that accounts for the largest tissue mass in the human body, is responsible for supporting posture, voluntary movement, guarding soft tissues and body openings, as well as regulating several metabolic and homoeostatic functions. Functional skeletal muscle not only contains myofibres and their progenitor cells but also requires their constant interaction with other cell types and tissues including, but not limited to, connective tissue, vasculature and motor neurons [1]. The hierarchical organisation of skeletal muscle (Figure 1a) consists of organised bundles of fascicles which in turn are composed of bundles of myofibres embedded within three layers of extracellular matrix (the endomysium, perimysium and epimysium) [2]. The importance of the interplay between different compartments of the skeletal muscle niche (Figure 1b) is exemplified on acute injury, when multiple mechanisms are initiated within the different compartments that eventually converge to activate tissue-resident muscle stem cells (MuSCs, also known as satellite cells). For instance, damaged blood vessels can release cytokines [3] or inflammatory cells [4] to support regeneration at an injury site.
Normal tissue function and repair/regeneration can be overcome in large acute muscle injuries as well as in chronic severe musculoskeletal disorders such as muscular dystrophy [5], where different components of the skeletal muscle tissue functional units and niche are compromised. Given ethical considerations and limited tissue availability, it is often difficult to study skeletal muscle developmental dynamics, regeneration and disease pathogenesis in human subjects or their biopsies. Although traditional cell culture and animal models have been used to elucidate some molecular aspects behind these processes, limitations in using different species [6] and systems lacking physiologically relevant extracellular cues [7] make it difficult to translate such findings to the human context. Bioengineering human models with higher fidelity to native skeletal muscle tissues can overcome these limitations and enable researchers to advance our fundamental understanding of the mechanistic processes behind muscle development and regeneration. Such insights can be further applied to disease modelling, biomarker detection, drug screening and regenerative medicine.
In this review, we will start with a brief overview of skeletal myogenic cell generation and differentiation followed by a discussion on recently developed three-dimensional (3D) platforms, developed with human biopsy-derived myoblasts (primary or immortalised) or pluripotent stem cells. We then conclude with our perspectives on the future of artificial skeletal muscle models by discussing methods to develop physiologically complex models able to deliver clinically relevant phenotypic readouts that can be used as outcome measures for therapy development. We will not highlight studies based on platforms using rodent myogenic cells, nor those involving top-down approaches such as tissue decellularization, for which we redirect the reader to recent comprehensive reviews [8,9].
Section snippets
Immortalising biopsy-derived skeletal myogenic cells
The ability to culture primary myogenic cells from human skeletal muscle biopsies ex vivo is crucial for modelling skeletal muscle function and disease [10,11]. However, the limited availability of patient tissue biopsies and restricted proliferative capacity of the extracted myoblasts make it difficult to use these cells extensively [12]. As a result, several immortalisation strategies have been applied to overcome Hayflick's limit while maintaining the myogenic differentiation potential of
Recapitulating 3D tissue complexity
Strategies to engineer 3D human skeletal muscles can be broadly classified into either 1) self-organised, organoid-like 3D cultures or 2) scaffold-based platforms. Recent notable studies using 3D culture platforms containing human myogenic cells are summarised in Table 2 and discussed in the following sections.
Future perspectives
The aphorism from the statistician George E. P. Box, ‘all models are wrong, but some are useful’, concisely summarises the current landscape of cellular modelling of skeletal muscle tissue development, differentiation and disease. Although none of the existing models discussed in this review fully recapitulate all aspects of the physiological skeletal muscle tissue niche, the ability to recreate at least some features has been invaluable to improve our understanding of skeletal muscle growth,
Conflict of interest statement
FST provides consulting services to Aleph Farms via UCL Consultants. The other authors do not declare conflict of interest.
Acknowledgements
This work was supported by the European Research Council (759108 – HISTOID) and the Francis Crick Institute, which receives its core funding from the Cancer Research UK, the UK Medical Research Council and the Welcome Trust (FC001002); Muscular Dystrophy UK (19GRO-PS48-0188; 17GRO-PS48-0093-1), the BBSRC and the NIHR (the views expressed are those of the authors and not necessarily those of the National Health Service, the NIHR or the Department of Health). Work on 3D human skeletal muscle
References (111)
- et al.
Muscular dystrophies
Lancet
(2019) - et al.
Decellularized tissues as platforms for in vitro modeling of healthy and diseased tissues
Acta Biomater
(2020) - et al.
Molecular basis of the myogenic profile of aged human skeletal muscle satellite cells during differentiation
Exp Gerontol
(2009) - et al.
Establishment of long-term myogenic cultures from patients with duchenne muscular dystrophy by retroviral transduction of a temperature-sensitive SV40 large T antigen
Exp Cell Res
(1996) - et al.
Reliable and versatile immortal muscle cell models from healthy and myotonic dystrophy type 1 primary human myoblasts
Exp Cell Res
(2016) Satellite cell of skeletal muscle fibers
J Biophys Biochem Cytol
(1961)- et al.
Transplantation of genetically corrected human iPSC-derived progenitors in mice with limb-girdle muscular dystrophy
Sci Transl Med
(2012) - et al.
A zebrafish embryo culture system defines factors that promote vertebrate myogenesis across species
Cell
(2013) - et al.
Three-dimensional human iPSC-derived artificial skeletal muscles model muscular dystrophies and enable multilineage tissue engineering
Cell Rep
(2018) - et al.
Matrix elasticity directs stem cell lineage specification
Cell
(2006)
Substrate elasticity regulates skeletal muscle stem cell self-renewal in culture
Science
Self-organizing 3D human trunk neuromuscular organoids
Cell Stem Cell
Bioengineered skeletal muscle as a model of muscle aging and regeneration
Tissue Eng
Neural cell integration into 3D bioprinted skeletal muscle constructs accelerates restoration of muscle function
Nat Commun
Bioengineered human skeletal muscle capable of functional regeneration
BMC Biol
Skeletal muscle constructs engineered from human embryonic stem cell derived myogenic progenitors exhibit enhanced contractile forces when differentiated in a medium containing EGM-2 supplements
Adv Biosys
Exercise mimetics and JAK inhibition attenuate IFN-γ–induced wasting in engineered human skeletal muscle
Sci Adv
A patient-derived iPSC model revealed oxidative stress increases facioscapulohumeral muscular dystrophy-causative DUX4
Hum Mol Genet
In vitro evaluation of exon skipping in disease-specific iPSC-derived myocytes
Efficient and reproducible myogenic differentiation from human iPS cells: prospects for modeling Miyoshi Myopathy in vitro
PloS One
Multilineage differentiation for formation of innervated skeletal muscle fibers from healthy and diseased human pluripotent stem cells
Cells
Vivo human somitogenesis guides somite development from hPSCs
Cell Rep
A novel protocol for directed differentiation of C9orf72-associated human induced pluripotent stem cells into contractile skeletal myotubes
STEM CELLS Transl Med
Differentiation of pluripotent stem cells to muscle fiber to model Duchenne muscular dystrophy
Nat Biotechnol
Derivation of myogenic progenitors directly from human pluripotent stem cells using a sphere-based culture
Stem Cells Transl Med
Skeletal muscle: a brief review of structure and function
Calcif Tissue Int
Skeletal muscle extracellular matrix – what do we know about its composition, regulation, and physiological roles? A narrative review
Front Physiol
Muscle satellite cells and endothelial cells: close neighbors and privileged partners
Mol Biol Cell
Macrophages provide a transient muscle stem cell niche via NAMPT secretion
Nature
Natural disease history of the D2-mdx mouse model for Duchenne muscular dystrophy
Faseb J
Modeling skeletal muscle laminopathies using human induced pluripotent stem cells carrying pathogenic LMNA mutations
Front Physiol
Decellularized tissue for muscle regeneration
Int J Mol Sci
Comparative profiling of skeletal muscle models reveals heterogeneity of transcriptome and metabolism
Am J Physiol Cell Physiol
Derivation and characterization of immortalized human muscle satellite cell clones from muscular dystrophy patients and healthy individuals
Cells
Cellular senescence in human myoblasts is overcome by human telomerase reverse transcriptase and cyclin-dependent kinase 4: consequences in aging muscle and therapeutic strategies for muscular dystrophies
Aging Cell
Lentivector-mediated transfer of Bmi-1 and telomerase in muscle satellite cells yields a duchenne myoblast cell line with long-term genotypic and phenotypic stability
Hum Gene Ther
Telomerase activity is sufficient to allow transformed cells to escape from crisis
Mol Cell Biol
CDK4 and cyclin D1 allow human myogenic cells to recapture growth property without compromising differentiation potential
Gene Ther
Skeletal muscle characteristics are preserved in hTERT/cdk4 human myogenic cell lines
Skeletal Muscle
Reversible immortalisation enables genetic correction of human muscle progenitors and engineering of next-generation human artificial chromosomes for Duchenne muscular dystrophy
EMBO Mol Med
Satellite cells delivered in their niche efficiently generate functional myotubes in three-dimensional cell culture
PloS One
A 3D culture model of innervated human skeletal muscle enables studies of the adult neuromuscular junction
eLife
Immortalized pathological human myoblasts: towards a universal tool for the study of neuromuscular disorders
Skeletal Muscle
Differentiation reveals latent features of aging and an energy barrier in murine myogenesis
Cell Rep
Two myogenic lineages within the developing somite
Development
Pax3/Pax7 mark a novel population of primitive myogenic cells during development
Gene Dev
A Pax3/Pax7-dependent population of skeletal muscle progenitor cells
Nature
MyoD or Myf-5 is required for the formation of skeletal muscle
Cell
In vivo satellite cell activation via Myf5 and MyoD in regenerating mouse skeletal muscle
J Cell Sci
Developmental myosins: expression patterns and functional significance
Skeletal Muscle
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These authors contributed equally to this work.