Muscle loss and impairment resulting from traumatic injury can be alleviated by therapies using muscle stem cells. However, collecting sufficient numbers of autologous myogenic stem cells and expanding them efficiently has been challenging. Here we show that myogenic stem cells (predominantly Pax7+ cells)—which were selectively expanded from readily obtainable dermal fibroblasts or skeletal muscle stem cells using a specific cocktail of small molecules and transplanted into muscle injuries in adult, aged or dystrophic mice—led to functional muscle regeneration in the three animal models. We also show that sustained release of the small-molecule cocktail in situ through polymer nanoparticles led to muscle repair by inducing robust activation and expansion of resident satellite cells. Chemically induced stem cell expansion in vitro and in situ may prove to be advantageous for stem cell therapies that aim to regenerate skeletal muscle and other tissues.
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npj Regenerative Medicine Open Access 08 August 2023
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Intramuscular delivery of neural crest stem cell spheroids enhances neuromuscular regeneration after denervation injury
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The main data supporting the results in this study are available within the paper and its Supplementary Information. Data for the microarray and scRNA-seq have been deposited in the NCBI Gene Expression Omnibus under accession numbers GSE158690 and GSE158691, respectively. All data generated in this study, including source data and the data used to make the figures, are available at Figshare (https://doi.org/10.6084/m9.figshare.13049690.v1).
The custom code used is available at GitHub (https://github.com/junrensia/Jun_et_al_Nature_BME_2020).
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We thank M. Conboy and V. Rezek for their technical assistance with the in vivo cell transplantation study and K. Heydari for his assistance with FACS. This work was supported in part by grants from UCLA Broad Stem Cell Research Center, the National Institute of Health (EB012240 HL121450 and R56DE029157 to S.L.), a fellowship from the Agency for Science, Technology and Research (to J. Sia), a fellowship from the National Institute of Arthritis and Musculoskeletal and Skin Diseases of the NIH under the Ruth L. Kirschstein National Research Service Award (T32AR059033 (to J. Soto) and the Medical Scientist Training Program at UCLA (NIH T32 GM008042 to L.K.L.). SEM was performed at the California NanoSystems Institute (CNSI) Electron Imaging Center for NanoMachines (EICN) Shared Resource Facility at UCLA. Confocal laser scanning microscopy was performed at the California NanoSystems Institute (CNSI) Advanced Light Microscopy/Spectroscopy Shared Resource Facility at UCLA. FACS was performed at the UCLA Jonsson Comprehensive Cancer Center (JCCC) and Center for AIDS Research Flow Cytometry Core Facility that is supported by National Institutes of Health awards P30 CA016042 and 5P30 AI028697, and by the JCCC, the UCLA AIDS Institute, the David Geffen School of Medicine at UCLA, the UCLA Chancellor’s Office and the UCLA Vice Chancellor’s Office of Research. Single-cell RNA sequencing was conducted at the UCLA Technology Center for Genomics and Bioinformatics.
The innovation related to this study has been filed for patent application by J.F., S.L. and J. Sia. (US Serial No. 30435411.2020).
Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Supplementary Figs. 1–16 and Table 1, and captions for Supplementary Videos 1–4.
Spontaneous beating after treatment with FR medium for 4 d.
Spontaneous beating after treatment with FR medium for 8 d.
Spontaneous beating after treatment with FR medium for 14 d.
Spontaneous beating after treatment with FR medium for 16 d.
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Fang, J., Sia, J., Soto, J. et al. Skeletal muscle regeneration via the chemical induction and expansion of myogenic stem cells in situ or in vitro. Nat Biomed Eng 5, 864–879 (2021). https://doi.org/10.1038/s41551-021-00696-y
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