Song Wenpeng, Wang Jian, Gong Shixin, Wang Xiaoyan, Xu Junji, Wu Ruiqing, Jiang Zongmin, Zhang Huiyuan, Wu Lida, Wang Yilong, Su Yingying, Wang Hao, Gu Yuchun
Department of Stomatology, Beijing Tiantan Hospital, Capital Medical University, Beijing, China.
Research and Development Department, Allife Medicine Inc., Beijing, China.
Theranostics. 2025 Jun 12;15(14):7127-7153. doi: 10.7150/thno.111198. eCollection 2025.
Pluripotent stem cells (PSCs) serve as a critical source of seed cells for regenerative therapies due to their unlimited proliferative capacity and ability to differentiate into all three germ layers. Despite their potential, the risk of teratoma formation caused by residual PSCs within differentiated cell populations poses a significant barrier to clinical applications. This study aims to develop a novel strategy to selectively remove residual PSCs while preserving the safety and functionality of PSC-derived differentiated cells (iDCs). The calcium- and magnesium-free balanced salt solution (BSS(Ca-Mg-)) was employed to selectively target PSCs in a co-culture system comprising PSCs and four types of iDCs. The effect of BSS(Ca-Mg-) treatment on teratoma formation was evaluated in immunodeficient mice following cell transplantation. Comparative analysis and gene knockdown experiments were conducted to explore the molecular mechanisms underlying the differential response of PSCs and iDCs to BSS(Ca-Mg-), focusing on FAK signaling and its interaction with OCT4 and ITGA6. The BSS(Ca-Mg-) treatment effectively induced the detachment of PSCs in the co-culture system without disrupting iDC adhesion. experiments confirmed that cells treated with BSS(Ca-Mg-) did not form teratomas upon implantation into immunodeficient mice. Mechanistic studies revealed that PSCs exhibit lower activation of FAK signaling compared to iDCs, contributing to their selective detachment. Additionally, OCT4 and ITGA6 were found to maintain each other's protein expression, forming a feedback loop that suppressed FAK signaling, while FAK suppression further enhanced OCT4 expression. The study presents a safe, effective, and cost-efficient method for the selective removal of residual PSCs. This approach enhances existing safety measures for iDC applications, improving the clinical feasibility of iDC-based cell therapies.
多能干细胞（PSCs）因其无限的增殖能力和分化为所有三个胚层的能力，成为再生疗法中种子细胞的关键来源。尽管具有潜力，但分化细胞群体中残留的PSCs导致畸胎瘤形成的风险，对临床应用构成了重大障碍。本研究旨在开发一种新策略，选择性去除残留的PSCs，同时保留PSCs来源的分化细胞（iDCs）的安全性和功能。在包含PSCs和四种类型iDCs的共培养系统中，使用无钙镁平衡盐溶液（BSS（Ca-Mg-））选择性靶向PSCs。细胞移植后，在免疫缺陷小鼠中评估BSS（Ca-Mg-）处理对畸胎瘤形成的影响。进行了比较分析和基因敲低实验，以探索PSCs和iDCs对BSS（Ca-Mg-）差异反应的分子机制，重点关注FAK信号及其与OCT4和ITGA6的相互作用。BSS（Ca-Mg-）处理有效地诱导了共培养系统中PSCs的脱离，而不破坏iDC的粘附。实验证实，用BSS（Ca-Mg-）处理的细胞植入免疫缺陷小鼠后不会形成畸胎瘤。机制研究表明，与iDCs相比，PSCs表现出较低的FAK信号激活，这有助于它们的选择性脱离。此外，发现OCT4和ITGA6相互维持蛋白表达，形成一个抑制FAK信号的反馈环，而FAK抑制进一步增强OCT4表达。该研究提出了一种安全、有效且经济高效的选择性去除残留PSCs的方法。这种方法增强了iDC应用的现有安全措施，提高了基于iDC的细胞疗法的临床可行性。
