Engineering Skeletal Muscle Grafts with PAX7::GFP-Sorted Human Pluripotent Stem Cell-Derived Myogenic Progenitors on Fibrin Microfiber Bundles for Tissue Regeneration.
Engineering Skeletal Muscle Grafts with PAX7::GFP-Sorted Human Pluripotent Stem Cell-Derived Myogenic Progenitors on Fibrin Microfiber Bundles for Tissue Regeneration.
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DOI：
10.3390/bioengineering9110693
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发表时间：
2022-11-15
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将人类多能干细胞衍生的肌源祖细胞 (hPDM) 与先进生物材料相结合的组织工程策略为工程 3D 骨骼肌移植物提供了有前途的工具，以模拟体外组织发育并促进体内肌肉再生。我们最近证明了（i）使用两种小分子的组合获得大量 hPDM 而不过度表达转基因的潜力，以及（ii）应用电纺纤维蛋白微纤维束在体积肌损失后恢复功能性骨骼肌。在这项研究中，我们的目的是证明纤维蛋白微纤维束提供的生物物理信号诱导 hPDM 形成工程化的人类骨骼肌移植物，其中含有表达结蛋白和肌球蛋白重链的多核肌管，并且这些移植物可以促进骨骼肌损伤后的再生。我们测试了遗传 PAX7 报告基因系 (PAX7::GFP)，以筛选出更同质的 hPDM 群体。 RNA 测序和基因集富集分析证实，PAX7::GFP 分选的 hPDM 表现出肌源性基因的高表达。我们通过使用免疫组织化学染色评估肌生成、植入和免疫原性，在体内骨骼肌缺陷内测试了源自 PAX7::GFP 分选的 hPDM 的工程人类骨骼肌移植物。 PAX7::GFP 分类组具有中等程度的高血管浸润，并且与胚胎肌球蛋白重链 (eMHC) 区域有更多的植入细胞关联，表明它们诱导了促再生微环境。这些发现证明了在纤维蛋白微纤维束上使用 PAX7::GFP 分选的 hPDM 的前景，并为改善细胞生物材料系统以刺激更强大的体内骨骼肌再生提供了一些见解。
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.
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影响因子：
8.8
作者：
Maffioletti SM;Sarcar S;Henderson ABH;Mannhardt I;Pinton L;Moyle LA;Steele-Stallard H;Cappellari O;Wells KE;Ferrari G;Mitchell JS;Tyzack GE;Kotiadis VN;Khedr M;Ragazzi M;Wang W;Duchen MR;Patani R;Zammit PS;Wells DJ;Eschenhagen T;Tedesco FS
通讯作者：
Tedesco FS
影响因子：
64.8
作者：
Cao, Junyue;Spielmann, Malte;Shendure, Jay
通讯作者：
Shendure, Jay
影响因子：
5.8
作者：
Liberzon, Arthur;Subramanian, Aravind;Mesirov, Jill P.
通讯作者：
Mesirov, Jill P.
DOI：
10.1093/bioinformatics/btu638
发表时间：
2015-01-15
期刊：
Bioinformatics (Oxford, England)
影响因子：
--
作者：
Anders S;Pyl PT;Huber W
通讯作者：
Huber W
DOI：
10.1016/j.bbrc.2019.06.082
发表时间：
2019-08-20
影响因子：
3.1
作者：
Guo, Yanheng;Gilbert-Honick, Jordana;Grayson, Warren L.
通讯作者：
Grayson, Warren L.