Selection against undifferentiated human embryonic stem cells by a cytotoxic antibody recognizing podocalyxin-like protein-1
Selection against undifferentiated human embryonic stem cells by a cytotoxic antibody recognizing podocalyxin-like protein-1
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DOI：
10.1634/stemcells.2007-0576
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发表时间：
2008-06-01
期刊：
影响因子：
5.2
通讯作者：
Yap, Miranda
中科院分区：
文献类型：
--
作者：
Choo, Andre B.;Tan, Heng Liang;Yap, Miranda
分化的人胚胎干细胞（hESC）在未来的治疗应用中存在因污染未分化的hESC而形成畸胎瘤的风险。我们制备了10种针对未分化hESC表面抗原的单克隆抗体（mab），对未分化但未分化的hESC表现出较强的反应性。单克隆抗体与小鼠成纤维细胞无交叉反应，对人胚胎癌细胞无反应。值得注意的是，其中一种抗体（mAb 84）对未分化的hESC和NCCIT细胞具有浓度依赖性、补体依赖性的细胞毒性。mAb 84在孵育30分钟内诱导未分化但未分化的hESC细胞死亡，免疫沉淀mAb-抗原复合物显示抗原为足alyxin样蛋白-1。重要的是，我们观察到，当hESC和NCCIT细胞在移植到严重联合免疫缺陷小鼠体内之前用mAb 84处理时，没有形成肿瘤。我们的数据表明，mAb 84可能有助于消除分化细胞群体中残留的hESC，用于临床应用。
Future therapeutic applications of differentiated human embryonic stem cells (hESC) carry a risk of teratoma formation by contaminating undifferentiated hESC. We generated 10 monoclonal antibodies (mAbs) against surface antigens of undifferentiated hESC, showing strong reactivity against undifferentiated, but not differentiated hESC. The mAbs did not cross react with mouse fibroblasts and showed weak to no reactivity against human embryonal carcinoma cells. Notably, one antibody (mAb 84) is cytotoxic to undifferentiated hESC and NCCIT cells in a concentration-dependent, complement-independent manner. mAb 84 induced cell death of undifferentiated, but not differentiated hESC within 30 minutes of incubation, and immunoprecipitation of the mAb-antigen complex revealed that the antigen is podocalyxin-like protein-1. Importantly, we observed absence of tumor formation when hESC and NCCIT cells were treated with mAb 84 prior to transplantation into severe combined immunodeficiency mice. Our data indicate that mAb 84 may be useful in eliminating residual hESC from differentiated cells populations for clinical applications.