Development of autologous tissue-engineered vascular constructs using vascular smooth muscle cells (VSMCs) derived from human induced pluripotent stem cells (iPSCs) holds great potential in treating patients with vascular disease. However, preclinical, large animal iPSC-based cellular and tissue models are required to evaluate safety and efficacy prior to clinical application. Herein, swine iPSC (siPSC) lines were established by introducing doxycycline-inducible reprogramming factors into fetal fibroblasts from a line of inbred Massachusetts General Hospital miniature swine that accept tissue and organ transplants without immunosuppression within the line. Highly enriched, functional VSMCs were derived from siPSCs based on addition of ascorbic acid and inactivation of reprogramming factor via doxycycline withdrawal. Moreover, siPSC-VSMCs seeded onto biodegradable polyglycolic acid (PGA) scaffolds readily formed vascular tissues, which were implanted subcutaneously into immunodeficient mice and showed further maturation revealed by expression of the mature VSMC marker, smooth muscle myosin heavy chain. Finally, using a robust cellular self-assembly approach, we developed 3D scaffold-free tissue rings from siPSC-VSMCs that showed comparable mechanical properties and contractile function to those developed from swine primary VSMCs. These engineered vascular constructs, prepared from doxycycline-inducible inbred siPSCs, offer new opportunities for preclinical investigation of autologous human iPSC-based vascular tissues for patient treatment.

中文翻译：

---

来自近交猪诱导多能干细胞的血管平滑肌细胞，用于血管组织工程。


使用源自人类诱导多能干细胞（iPSC）的血管平滑肌细胞（VSMC）开发自体组织工程血管结构在治疗血管疾病患者方面具有巨大潜力。然而，在临床应用之前，需要临床前、基于大型动物 iPSC 的细胞和组织模型来评估安全性和有效性。在此，通过将多西环素诱导重编程因子引入马萨诸塞州总医院近交系小型猪的胎儿成纤维细胞中，建立了猪 iPSC (siPSC) 系，这些近交系小型猪接受组织和器官移植，且系内没有免疫抑制。通过添加抗坏血酸并通过强力霉素撤除使重编程因子失活，从 siPSC 中衍生出高度富集的功能性 VSMC。此外，接种到可生物降解的聚乙醇酸（PGA）支架上的siPSC-VSMC很容易形成血管组织，将其皮下植入免疫缺陷小鼠体内，并通过成熟VSMC标记物平滑肌肌球蛋白重链的表达显示进一步成熟。最后，使用强大的细胞自组装方法，我们从 siPSC-VSMC 开发了 3D 无支架组织环，其机械性能和收缩功能与从猪原代 VSMC 开发的组织环相当。这些工程化血管结构由强力霉素诱导的近交系 siPSC 制备而成，为用于患者治疗的自体人 iPSC 血管组织的临床前研究提供了新的机会。