Department of Genetics, Silberman Institute of Life Sciences, The Hebrew University, Jerusalem 91904, Israel.
Nat Commun. 2013;4:1992. doi: 10.1038/ncomms2992.
The tumorigenicity of human pluripotent stem cells is a major safety concern for their application in regenerative medicine. Here we identify the tight-junction protein Claudin-6 as a cell-surface-specific marker of human pluripotent stem cells that can be used to selectively remove Claudin-6-positive cells from mixed cultures. We show that Claudin-6 is absent in adult tissues but highly expressed in undifferentiated cells, where it is dispensable for human pluripotent stem cell survival and self-renewal. We use three different strategies to remove Claudin-6-positive cells from mixed cell populations: an antibody against Claudin-6; a cytotoxin-conjugated antibody that selectively targets undifferentiated cells; and Clostridium perfringens enterotoxin, a toxin that binds several Claudins, including Claudin-6, and efficiently kills undifferentiated cells, thus eliminating the tumorigenic potential of human pluripotent stem cell-containing cultures. This work provides a proof of concept for the use of Claudin-6 to eliminate residual undifferentiated human pluripotent stem cells from culture, highlighting a strategy that may increase the safety of human pluripotent stem cell-based cell therapies.
人多能干细胞的致瘤性是其在再生医学中应用的主要安全关注点。在这里，我们确定紧密连接蛋白 Claudin-6 是人多能干细胞的表面特异性标记物，可用于从混合培养物中选择性去除 Claudin-6 阳性细胞。我们表明 Claudin-6 在成年组织中不存在，但在未分化细胞中高度表达，在那里它对于人多能干细胞的存活和自我更新是可有可无的。我们使用三种不同的策略从混合细胞群中去除 Claudin-6 阳性细胞：抗 Claudin-6 抗体；一种针对未分化细胞的细胞毒素缀合抗体；以及产气荚膜梭菌肠毒素，一种与包括 Claudin-6 在内的几种 Claudin 结合的毒素，可有效杀死未分化细胞，从而消除含有人多能干细胞的培养物的致瘤潜能。这项工作为使用 Claudin-6 从培养物中去除残留的未分化人多能干细胞提供了概念验证，突出了一种可能提高基于人多能干细胞的细胞治疗安全性的策略。
