Department of Medicine, University of Minnesota, Minneapolis, MN 55455, USA.
Stem Cell Resources and the Wisconsin National Primate Research Center, University of Wisconsin, Madison, WI 53715, USA.
Cells. 2023 Apr 13;12(8):1147. doi: 10.3390/cells12081147.
Pluripotent stem (PS) cells enable the scalable production of tissue-specific derivatives with therapeutic potential for various clinical applications, including muscular dystrophies. Given the similarity to human counterparts, the non-human primate (NHP) is an ideal preclinical model to evaluate several questions, including delivery, biodistribution, and immune response. While the generation of human-induced PS (iPS)-cell-derived myogenic progenitors is well established, there have been no data for NHP counterparts, probably due to the lack of an efficient system to differentiate NHP iPS cells towards the skeletal muscle lineage. Here, we report the generation of three independent Macaca fascicularis iPS cell lines and their myogenic differentiation using PAX7 conditional expression. The whole-transcriptome analysis confirmed the successful sequential induction of mesoderm, paraxial mesoderm, and myogenic lineages. NHP myogenic progenitors efficiently gave rise to myotubes under appropriate in vitro differentiation conditions and engrafted in vivo into the TA muscles of NSG and FKRP-NSG mice. Lastly, we explored the preclinical potential of these NHP myogenic progenitors in a single wild-type NHP recipient, demonstrating engraftment and characterizing the interaction with the host immune response. These studies establish an NHP model system through which iPS-cell-derived myogenic progenitors can be studied.
多能干细胞（PS）能够规模化生成具有治疗潜力的组织特异性衍生物，可应用于多种临床疾病，包括肌肉萎缩症。鉴于与人类的相似性，非人类灵长类动物（NHP）是评估包括递送、生物分布和免疫反应在内的许多问题的理想临床前模型。虽然已经建立了人类诱导多能干细胞（iPS）衍生的成肌祖细胞的生成方法，但针对 NHP 对应物却没有数据，这可能是由于缺乏有效的系统来将 NHP iPS 细胞分化为骨骼肌谱系。在这里，我们报告了三个独立的食蟹猴 iPS 细胞系的生成，并使用 PAX7 条件表达对其进行了成肌分化。全转录组分析证实了中胚层、轴旁中胚层和肌源性谱系的成功连续诱导。在适当的体外分化条件下，NHP 成肌祖细胞能够有效地分化为肌管，并在体内植入 NSG 和 FKRP-NSG 小鼠的 TA 肌肉中。最后，我们在单个野生型 NHP 受体内探索了这些 NHP 成肌祖细胞的临床前潜力，证明了其植入和与宿主免疫反应的相互作用。这些研究建立了一个 NHP 模型系统，可以通过该系统研究 iPS 细胞衍生的成肌祖细胞。
