Standardization of the teratoma assay for analysis of pluripotency of human ES cells and biosafety of their differentiated progeny.
Standardization of the teratoma assay for analysis of pluripotency of human ES cells and biosafety of their differentiated progeny.
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DOI：
10.1371/journal.pone.0045532
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Reubinoff BE
中科院分区：
文献类型：
--
作者：
Gropp M;Shilo V;Vainer G;Gov M;Gil Y;Khaner H;Matzrafi L;Idelson M;Kopolovic J;Zak NB;Reubinoff BE
畸胎瘤肿瘤形成是确定人多能干细胞多能性的基本标准。然而，目前还没有一致的协议评估畸胎瘤形成的能力。在这里，我们提出了一个畸胎瘤检测，是基于皮下共移植的定义数量的未分化的人胚胎干细胞（hESC）与有丝分裂失活的饲养细胞和Matrigel到免疫缺陷小鼠的详细特征。当移植100，000个hESC时，该测定具有高度可重复性和100%的效率。它是敏感的，促进畸胎瘤形成后，移植100 hESC，虽然需要更多的动物和更长的随访。该测定可以检测分化的hESC群体中残留的畸胎瘤形成细胞，但在分化细胞存在的情况下，其灵敏度降低。我们的数据奠定了基础，标准化的畸胎瘤测定多能性分析。该测定也可用于多能干细胞衍生的分化后代的生物安全性分析。
Teratoma tumor formation is an essential criterion in determining the pluripotency of human pluripotent stem cells. However, currently there is no consistent protocol for assessment of teratoma forming ability. Here we present detailed characterization of a teratoma assay that is based on subcutaneous co-transplantation of defined numbers of undifferentiated human embryonic stem cells (hESCs) with mitotically inactivated feeder cells and Matrigel into immunodeficient mice. The assay was highly reproducible and 100% efficient when 100,000 hESCs were transplanted. It was sensitive, promoting teratoma formation after transplantation of 100 hESCs, though larger numbers of animals and longer follow-up were required. The assay could detect residual teratoma forming cells within differentiated hESC populations however its sensitivity was decreased in the presence of differentiated cells. Our data lay the foundation, for standardization of a teratoma assay for pluripotency analysis. The assay can also be used for bio-safety analysis of pluripotent stem cell-derived differentiated progeny.
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影响因子：
56.9
作者：
Thomson, JA;Itskovitz-Eldor, J;Jones, JM
通讯作者：
Jones, JM
影响因子：
4
作者：
Shih, Chu-Chih;Forman, Stephen J.;Slovak, Marilyn
通讯作者：
Slovak, Marilyn
影响因子：
46.9
作者：
Spits, Claudia;Mateizel, Ileana;Sermon, Karen
通讯作者：
Sermon, Karen
影响因子：
5.2
作者：
Drukker, Micha;Katchman, Helena;Benvenisty, Nissim
通讯作者：
Benvenisty, Nissim
影响因子：
46.9
作者：
Werbowetski-Ogilvie, Tamra E.;Bosse, Marc;Bhatia, Mickie
通讯作者：
Bhatia, Mickie