Lillehei Heart Institute, University of Minnesota, Minneapolis, MN, USA.
School of Kinesiology, University of Minnesota, Minneapolis, MN, USA.
Nat Biomed Eng. 2021 Aug;5(8):805-814. doi: 10.1038/s41551-021-00693-1. Epub 2021 Mar 29.
Because post-mortem human skeletal muscle is not viable, autologous muscle grafts are typically required in tissue reconstruction after muscle loss due to disease or injury. However, the use of autologous tissue often leads to donor-site morbidity. Here, we show that intraspecies and interspecies chimaeric pig embryos lacking native skeletal muscle can be produced by deleting the MYF5, MYOD and MYF6 genes in the embryos via CRISPR, followed by somatic-cell nuclear transfer and the delivery of exogenous cells (porcine blastomeres or human induced pluripotent stem cells) via blastocyst complementation. The generated intraspecies chimaeras were viable and displayed normal histology, morphology and function. Human:pig chimaeras generated with TP53-null human induced pluripotent stem cells led to higher chimaerism efficiency, with embryos collected at embryonic days 20 and 27 containing humanized muscle, as confirmed by immunohistochemical and molecular analyses. Human:pig chimaeras may facilitate the production of exogenic organs for research and xenotransplantation.
由于死后的人体骨骼肌无法存活，因此在疾病或损伤导致肌肉丧失后，组织重建通常需要使用自体肌肉移植物。然而，自体组织的使用往往会导致供体部位的发病率。在这里，我们通过 CRISPR 技术在胚胎中删除 MYF5、MYOD 和 MYF6 基因，然后进行体细胞核移植，并通过胚胎互补将外源性细胞（猪卵裂球或人诱导多能干细胞）递送至胚胎，从而展示了可以生成缺乏内源性骨骼肌的同种和异种嵌合猪胚胎。生成的同种嵌合体具有活力，并表现出正常的组织学、形态和功能。使用 TP53 缺失的人诱导多能干细胞生成的人-猪嵌合体导致更高的嵌合效率，通过免疫组织化学和分子分析证实，在胚胎第 20 天和第 27 天收集的胚胎中含有人源化的肌肉。人-猪嵌合体可能有助于为研究和异种移植生产外源器官。
