Introduction

Tumourigenesis attributed to residual undifferentiated cells in a graft is considered to be a significant issue in cell therapy using human pluripotent stem cells. To ensure the safety of regenerative medicine derived from pluripotent stem cells, residual undifferentiated cells must be eliminated in the manufacturing process. We previously described the lectin probe rBC2LCN, which binds harmlessly and specifically to the cell surface of human pluripotent stem cells. We report here a technique using rBC2LCN to remove pluripotent cells from a heterogenous population to reduce the chance of teratoma formation.

Methods

We demonstrate a method for separating residual tumourigenic cells using rBC2LCN-bound magnetic beads. This technology is a novel use of their previous discovery that rBC2LCN is a lectin that selectively binds to pluripotent cells. We optimize and validate a method to remove hPSCs from a mixture with human fibroblasts using rBC2LCN-conjugated magnetic beads.

Results

Cells with the potential to form teratoma could be effectively eliminated from a heterogeneous cell population with biotin-labelled rBC2LCN and streptavidin-bound magnetic beads. The efficiency was measured by FACS, ddPCR, and animal transplantation, suggesting that magnetic cell separation using rBC2LCN is quite efficient for eliminating hPSCs from mixed cell populations.

Conclusions

The removal of residual tumourigenic cells based on rBC2LCN could be a practical option for laboratory use and industrialisation of regenerative medicine using human pluripotent stem cells.

中文翻译：

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一种使用 rBC2LCN 凝集素去除致瘤多能干细胞的技术。

 介绍

由移植物中残留的未分化细胞引起的肿瘤发生被认为是使用人类多能干细胞的细胞治疗中的一个重要问题。为了确保多能干细胞再生医学的安全性，在制造过程中必须消除残留的未分化细胞。我们之前描述了凝集素探针 rBC2LCN，它能无害且特异性地结合到人类多能干细胞的细胞表面。我们在此报告了一种使用 rBC2LCN 从异质群体中去除多能细胞的技术，以减少畸胎瘤形成的机会。

 方法

我们展示了一种使用 rBC2LCN 结合磁珠分离残留致瘤细胞的方法。这项技术是他们之前发现的新用途，即 rBC2LCN 是一种选择性结合多能细胞的凝集素。我们优化并验证了一种使用 rBC2LCN 缀合磁珠从与人成纤维细胞的混合物中去除 hPSC 的方法。

 结果

使用生物素标记的 rBC2LCN 和链霉亲和素结合的磁珠，可以有效地从异质细胞群中消除可能形成畸胎瘤的细胞。通过 FACS、ddPCR 和动物移植测量效率，表明使用 rBC2LCN 的磁性细胞分离对于从混合细胞群中消除 hPSC 非常有效。

 结论

基于rBC2LCN去除残留致瘤细胞可能是实验室使用和利用人类多能干细胞进行再生医学工业化的实用选择。