Engineering Skeletal Muscle Grafts with PAX7::GFP-Sorted Human Pluripotent Stem Cell-Derived Myogenic Progenitors on Fibrin Microfiber Bundles for Tissue Regeneration
Tissue engineering strategies that combine human pluripotent stem cell-derived myogenic progenitors (hPDMs) with advanced biomaterials provide promising tools for engineering 3D skeletal muscle grafts to model tissue development in vitro and promote muscle regeneration in vivo. We recently demonstra...
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| Format: | Article |
| Language: | English |
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MDPI AG
2022-11-01
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| Series: | Bioengineering |
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| Online Access: | https://www.mdpi.com/2306-5354/9/11/693 |
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| author | Sarah M. Somers Jordana Gilbert-Honick In Young Choi Emily K. W. Lo HoTae Lim Shaquielle Dias Kathryn R. Wagner Hai-Quan Mao Patrick Cahan Gabsang Lee Warren L. Grayson |
| author_facet | Sarah M. Somers Jordana Gilbert-Honick In Young Choi Emily K. W. Lo HoTae Lim Shaquielle Dias Kathryn R. Wagner Hai-Quan Mao Patrick Cahan Gabsang Lee Warren L. Grayson |
| author_sort | Sarah M. Somers |
| collection | DOAJ |
| description | Tissue engineering strategies that combine human pluripotent stem cell-derived myogenic progenitors (hPDMs) with advanced biomaterials provide promising tools for engineering 3D skeletal muscle grafts to model tissue development in vitro and promote muscle regeneration in vivo. We recently demonstrated (i) the potential for obtaining large numbers of hPDMs using a combination of two small molecules without the overexpression of transgenes and (ii) the application of electrospun fibrin microfiber bundles for functional skeletal muscle restoration following volumetric muscle loss. In this study, we aimed to demonstrate that the biophysical cues provided by the fibrin microfiber bundles induce hPDMs to form engineered human skeletal muscle grafts containing multinucleated myotubes that express desmin and myosin heavy chains and that these grafts could promote regeneration following skeletal muscle injuries. We tested a genetic PAX7 reporter line (PAX7::GFP) to sort for more homogenous populations of hPDMs. RNA sequencing and gene set enrichment analyses confirmed that PAX7::GFP-sorted hPDMs exhibited high expression of myogenic genes. We tested engineered human skeletal muscle grafts derived from PAX7::GFP-sorted hPDMs within in vivo skeletal muscle defects by assessing myogenesis, engraftment and immunogenicity using immunohistochemical staining. The PAX7::GFP-sorted groups had moderately high vascular infiltration and more implanted cell association with embryonic myosin heavy chain (eMHC) regions, suggesting they induced pro-regenerative microenvironments. These findings demonstrated the promise for the use of PAX7::GFP-sorted hPDMs on fibrin microfiber bundles and provided some insights for improving the cell–biomaterial system to stimulate more robust in vivo skeletal muscle regeneration. |
| first_indexed | 2024-03-09T18:28:35Z |
| format | Article |
| id | doaj.art-a6b89d7824424fbe817383c8b7bdca00 |
| institution | Directory Open Access Journal |
| issn | 2306-5354 |
| language | English |
| last_indexed | 2024-03-09T18:28:35Z |
| publishDate | 2022-11-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Bioengineering |
| spelling | doaj.art-a6b89d7824424fbe817383c8b7bdca002023-11-24T07:44:34ZengMDPI AGBioengineering2306-53542022-11-0191169310.3390/bioengineering9110693Engineering Skeletal Muscle Grafts with PAX7::GFP-Sorted Human Pluripotent Stem Cell-Derived Myogenic Progenitors on Fibrin Microfiber Bundles for Tissue RegenerationSarah M. Somers0Jordana Gilbert-Honick1In Young Choi2Emily K. W. Lo3HoTae Lim4Shaquielle Dias5Kathryn R. Wagner6Hai-Quan Mao7Patrick Cahan8Gabsang Lee9Warren L. Grayson10Translational Tissue Engineering Center, Johns Hopkins University School of Medicine, Baltimore, MD 21231, USATranslational Tissue Engineering Center, Johns Hopkins University School of Medicine, Baltimore, MD 21231, USAThe Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USADepartment of Biomedical Engineering, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USAThe Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USATranslational Tissue Engineering Center, Johns Hopkins University School of Medicine, Baltimore, MD 21231, USAThe Hugo W. Moser Research Institute, Kennedy Krieger Institute, Baltimore, MD 21205, USATranslational Tissue Engineering Center, Johns Hopkins University School of Medicine, Baltimore, MD 21231, USADepartment of Biomedical Engineering, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USAThe Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USATranslational Tissue Engineering Center, Johns Hopkins University School of Medicine, Baltimore, MD 21231, USATissue engineering strategies that combine human pluripotent stem cell-derived myogenic progenitors (hPDMs) with advanced biomaterials provide promising tools for engineering 3D skeletal muscle grafts to model tissue development in vitro and promote muscle regeneration in vivo. We recently demonstrated (i) the potential for obtaining large numbers of hPDMs using a combination of two small molecules without the overexpression of transgenes and (ii) the application of electrospun fibrin microfiber bundles for functional skeletal muscle restoration following volumetric muscle loss. In this study, we aimed to demonstrate that the biophysical cues provided by the fibrin microfiber bundles induce hPDMs to form engineered human skeletal muscle grafts containing multinucleated myotubes that express desmin and myosin heavy chains and that these grafts could promote regeneration following skeletal muscle injuries. We tested a genetic PAX7 reporter line (PAX7::GFP) to sort for more homogenous populations of hPDMs. RNA sequencing and gene set enrichment analyses confirmed that PAX7::GFP-sorted hPDMs exhibited high expression of myogenic genes. We tested engineered human skeletal muscle grafts derived from PAX7::GFP-sorted hPDMs within in vivo skeletal muscle defects by assessing myogenesis, engraftment and immunogenicity using immunohistochemical staining. The PAX7::GFP-sorted groups had moderately high vascular infiltration and more implanted cell association with embryonic myosin heavy chain (eMHC) regions, suggesting they induced pro-regenerative microenvironments. These findings demonstrated the promise for the use of PAX7::GFP-sorted hPDMs on fibrin microfiber bundles and provided some insights for improving the cell–biomaterial system to stimulate more robust in vivo skeletal muscle regeneration.https://www.mdpi.com/2306-5354/9/11/693human pluripotent stem cells (hPSCs)hPSC-derived myogenic progenitors (hPDMs) PAX7skeletal muscle tissue engineeringskeletal muscle injury and regenerationfibrin microfiber bundles |
| spellingShingle | Sarah M. Somers Jordana Gilbert-Honick In Young Choi Emily K. W. Lo HoTae Lim Shaquielle Dias Kathryn R. Wagner Hai-Quan Mao Patrick Cahan Gabsang Lee Warren L. Grayson Engineering Skeletal Muscle Grafts with PAX7::GFP-Sorted Human Pluripotent Stem Cell-Derived Myogenic Progenitors on Fibrin Microfiber Bundles for Tissue Regeneration Bioengineering human pluripotent stem cells (hPSCs) hPSC-derived myogenic progenitors (hPDMs) PAX7 skeletal muscle tissue engineering skeletal muscle injury and regeneration fibrin microfiber bundles |
| title | Engineering Skeletal Muscle Grafts with PAX7::GFP-Sorted Human Pluripotent Stem Cell-Derived Myogenic Progenitors on Fibrin Microfiber Bundles for Tissue Regeneration |
| title_full | Engineering Skeletal Muscle Grafts with PAX7::GFP-Sorted Human Pluripotent Stem Cell-Derived Myogenic Progenitors on Fibrin Microfiber Bundles for Tissue Regeneration |
| title_fullStr | Engineering Skeletal Muscle Grafts with PAX7::GFP-Sorted Human Pluripotent Stem Cell-Derived Myogenic Progenitors on Fibrin Microfiber Bundles for Tissue Regeneration |
| title_full_unstemmed | Engineering Skeletal Muscle Grafts with PAX7::GFP-Sorted Human Pluripotent Stem Cell-Derived Myogenic Progenitors on Fibrin Microfiber Bundles for Tissue Regeneration |
| title_short | Engineering Skeletal Muscle Grafts with PAX7::GFP-Sorted Human Pluripotent Stem Cell-Derived Myogenic Progenitors on Fibrin Microfiber Bundles for Tissue Regeneration |
| title_sort | engineering skeletal muscle grafts with pax7 gfp sorted human pluripotent stem cell derived myogenic progenitors on fibrin microfiber bundles for tissue regeneration |
| topic | human pluripotent stem cells (hPSCs) hPSC-derived myogenic progenitors (hPDMs) PAX7 skeletal muscle tissue engineering skeletal muscle injury and regeneration fibrin microfiber bundles |
| url | https://www.mdpi.com/2306-5354/9/11/693 |
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