Tang Peng, Wei Fuxiang, Qiao Weihua, Chen Xing, Ji Chenyang, Yang Wanzhi, Zhang Xinyu, Chen Sihan, Wu Yanyan, Jiang Mingxing, Ma Chenyu, Shen Weiqiang, Dong Qi, Cao Hong, Xie Minghui, Cai Ziwen, Xu Li, Shi Jiawei, Dong Nianguo, Chen Junwei, Wang Ning
Key Laboratory of Molecular Biophysics of the Ministry of Education, Hubei Bioinformatics and Molecular Imaging Key Laboratory, Laboratory for Cellular Biomechanics and Regenerative Medicine, Department of Biomedical Engineering, College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan, Hubei, 430074, China.
Department of Cardiovascular Surgery, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430022, China.
Bioact Mater. 2024 Nov 23;45:181-200. doi: 10.1016/j.bioactmat.2024.11.018. eCollection 2025 Mar.
The technology of induced pluripotent stem cells (iPSCs) has enabled the conversion of somatic cells into primitive undifferentiated cells via reprogramming. This approach provides possibilities for cell replacement therapies and drug screening, but the potential risk of tumorigenesis hampers its further development and application. How to generate differentiated cells such as valvular endothelial cells (VECs) has remained a major challenge. Utilizing a combinatorial strategy of selective soluble chemicals, cytokines and substrate stiffness modulation, mouse embryonic fibroblasts are directly and efficiently transdifferentiated into induced aortic endothelial cell-like cells (iAECs), or human primary adult fibroblasts are transdifferentiated into induced valvular endothelial cell-like cells (hiVECs), without expressing pluripotency stem cell markers. These iAECs and hiVECs express VEC-associated genes and proteins and VEC-specific marker NFATC1 and are functional in culture and on decellularized porcine aortic valves, like mouse aortic endothelial cells or human primary aortic valvular endothelial cells. The iAECs and hiVECs seeded on decellularized porcine aortic valves stay intact and express VEC-associated proteins for 60 days after grafting into abdominal aorta of immune-compromised rats. In contrast, induced pluripotent stem cells (iPSCs) are less efficient in differentiating into VEC-like cells and pluripotency marker Nanog is expressed in a small subpopulation of iPSC-derived VEC-like cells that generate teratomas in SCID mice whereas hiVECs derived from transdifferentiation do not generate teratomas . Our findings highlight an approach to efficiently convert fibroblasts into iAECs and hiVECs and seed them onto decellularized aortic valves for safely generating autologous tissue-engineered aortic valves without using viruses or first reprogramming the cells into pluripotent stem cells.
诱导多能干细胞（iPSC）技术能够通过重编程将体细胞转化为原始未分化细胞。这种方法为细胞替代疗法和药物筛选提供了可能性，但肿瘤发生的潜在风险阻碍了其进一步发展和应用。如何生成诸如瓣膜内皮细胞（VEC）等分化细胞仍然是一个重大挑战。利用选择性可溶性化学物质、细胞因子和底物硬度调节的组合策略，小鼠胚胎成纤维细胞可直接且高效地转分化为诱导性主动脉内皮样细胞（iAEC），或者人类原代成纤维细胞可转分化为诱导性瓣膜内皮样细胞（hiVEC），且不表达多能干细胞标志物。这些iAEC和hiVEC表达与VEC相关的基因和蛋白质以及VEC特异性标志物NFATC1，并且在培养物中和脱细胞猪主动脉瓣膜上具有功能，类似于小鼠主动脉内皮细胞或人类原代主动脉瓣膜内皮细胞。接种在脱细胞猪主动脉瓣膜上的iAEC和hiVEC在移植到免疫缺陷大鼠的腹主动脉后60天内保持完整并表达与VEC相关的蛋白质。相比之下，诱导多能干细胞（iPSC）在分化为VEC样细胞方面效率较低，并且多能性标志物Nanog在一小部分iPSC衍生的VEC样细胞中表达，这些细胞在SCID小鼠中会形成畸胎瘤，而转分化衍生的hiVEC则不会形成畸胎瘤。我们的研究结果突出了一种方法，即有效地将成纤维细胞转化为iAEC和hiVEC，并将它们接种到脱细胞主动脉瓣膜上，以安全地生成自体组织工程主动脉瓣膜，而无需使用病毒或将细胞先重编程为多能干细胞。
