Cargando…

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...

Descripción completa

Detalles Bibliográficos
Autores principales: Somers, Sarah M., Gilbert-Honick, Jordana, Choi, In Young, K. W. Lo, Emily, Lim, HoTae, Dias, Shaquielle, Wagner, Kathryn R., Mao, Hai-Quan, Cahan, Patrick, Lee, Gabsang, Grayson, Warren L.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9687588/
https://www.ncbi.nlm.nih.gov/pubmed/36421094
http://dx.doi.org/10.3390/bioengineering9110693
_version_ 1784836043816566784
author Somers, Sarah M.
Gilbert-Honick, Jordana
Choi, In Young
K. W. Lo, Emily
Lim, HoTae
Dias, Shaquielle
Wagner, Kathryn R.
Mao, Hai-Quan
Cahan, Patrick
Lee, Gabsang
Grayson, Warren L.
author_facet Somers, Sarah M.
Gilbert-Honick, Jordana
Choi, In Young
K. W. Lo, Emily
Lim, HoTae
Dias, Shaquielle
Wagner, Kathryn R.
Mao, Hai-Quan
Cahan, Patrick
Lee, Gabsang
Grayson, Warren L.
author_sort Somers, Sarah M.
collection PubMed
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.
format Online
Article
Text
id pubmed-9687588
institution National Center for Biotechnology Information
language English
publishDate 2022
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-96875882022-11-25 Engineering Skeletal Muscle Grafts with PAX7::GFP-Sorted Human Pluripotent Stem Cell-Derived Myogenic Progenitors on Fibrin Microfiber Bundles for Tissue Regeneration Somers, Sarah M. Gilbert-Honick, Jordana Choi, In Young K. W. Lo, Emily Lim, HoTae Dias, Shaquielle Wagner, Kathryn R. Mao, Hai-Quan Cahan, Patrick Lee, Gabsang Grayson, Warren L. Bioengineering (Basel) Article 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. MDPI 2022-11-15 /pmc/articles/PMC9687588/ /pubmed/36421094 http://dx.doi.org/10.3390/bioengineering9110693 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Somers, Sarah M.
Gilbert-Honick, Jordana
Choi, In Young
K. W. Lo, Emily
Lim, HoTae
Dias, Shaquielle
Wagner, Kathryn R.
Mao, Hai-Quan
Cahan, Patrick
Lee, Gabsang
Grayson, Warren L.
Engineering Skeletal Muscle Grafts with PAX7::GFP-Sorted Human Pluripotent Stem Cell-Derived Myogenic Progenitors on Fibrin Microfiber Bundles for Tissue Regeneration
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 Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9687588/
https://www.ncbi.nlm.nih.gov/pubmed/36421094
http://dx.doi.org/10.3390/bioengineering9110693
work_keys_str_mv AT somerssarahm engineeringskeletalmusclegraftswithpax7gfpsortedhumanpluripotentstemcellderivedmyogenicprogenitorsonfibrinmicrofiberbundlesfortissueregeneration
AT gilberthonickjordana engineeringskeletalmusclegraftswithpax7gfpsortedhumanpluripotentstemcellderivedmyogenicprogenitorsonfibrinmicrofiberbundlesfortissueregeneration
AT choiinyoung engineeringskeletalmusclegraftswithpax7gfpsortedhumanpluripotentstemcellderivedmyogenicprogenitorsonfibrinmicrofiberbundlesfortissueregeneration
AT kwloemily engineeringskeletalmusclegraftswithpax7gfpsortedhumanpluripotentstemcellderivedmyogenicprogenitorsonfibrinmicrofiberbundlesfortissueregeneration
AT limhotae engineeringskeletalmusclegraftswithpax7gfpsortedhumanpluripotentstemcellderivedmyogenicprogenitorsonfibrinmicrofiberbundlesfortissueregeneration
AT diasshaquielle engineeringskeletalmusclegraftswithpax7gfpsortedhumanpluripotentstemcellderivedmyogenicprogenitorsonfibrinmicrofiberbundlesfortissueregeneration
AT wagnerkathrynr engineeringskeletalmusclegraftswithpax7gfpsortedhumanpluripotentstemcellderivedmyogenicprogenitorsonfibrinmicrofiberbundlesfortissueregeneration
AT maohaiquan engineeringskeletalmusclegraftswithpax7gfpsortedhumanpluripotentstemcellderivedmyogenicprogenitorsonfibrinmicrofiberbundlesfortissueregeneration
AT cahanpatrick engineeringskeletalmusclegraftswithpax7gfpsortedhumanpluripotentstemcellderivedmyogenicprogenitorsonfibrinmicrofiberbundlesfortissueregeneration
AT leegabsang engineeringskeletalmusclegraftswithpax7gfpsortedhumanpluripotentstemcellderivedmyogenicprogenitorsonfibrinmicrofiberbundlesfortissueregeneration
AT graysonwarrenl engineeringskeletalmusclegraftswithpax7gfpsortedhumanpluripotentstemcellderivedmyogenicprogenitorsonfibrinmicrofiberbundlesfortissueregeneration