Assessment of established techniques to determine developmental and malignant potential of human pluripotent stem cells
Thomas F. AllisonPeter W. AndrewsYishai AviorIvana BarbaricNissim BenvenistyChristoph BockJennifer BrehmOliver BrüstleIvan DamjanovAndrew G. ElefantyDaniel FelknerPaul J. GokhaleFlorian HalbritterLyn HealyXiaoming HuBarbara B. KnowlesJeanne F. LoringTenneille E. LudwigRobyn MayberrySuzanne J. MicallefJameelah Sheik MohamedFrank MüllerChristine L. MummeryNorio NakatsujiElizabeth S. NgSteve OhOrla O’SheaMartín F. PeraBenjamin ReubinoffPaul RobsonJanet RossantBernhard SchuldtDavor SolterKoula SourrisGlyn StaceyEdouard G. StanleyHirofumi SuemoriKazutoshi TakahashiShinya Yamanaka
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Abstract The International Stem Cell Initiative compared several commonly used approaches to assess human pluripotent stem cells (PSC). PluriTest predicts pluripotency through bioinformatic analysis of the transcriptomes of undifferentiated cells, whereas, embryoid body (EB) formation in vitro and teratoma formation in vivo provide direct tests of differentiation. Here we report that EB assays, analyzed after differentiation under neutral conditions and under conditions promoting differentiation to ectoderm, mesoderm, or endoderm lineages, are sufficient to assess the differentiation potential of PSCs. However, teratoma analysis by histologic examination and by TeratoScore, which estimates differential gene expression in each tumor, not only measures differentiation but also allows insight into a PSC’s malignant potential. Each of the assays can be used to predict pluripotent differentiation potential but, at this stage of assay development, only the teratoma assay provides an assessment of pluripotency and malignant potential, which are both relevant to the pre-clinical safety assessment of PSCs.Keywords:
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During embryonic development, primitive ectoderm forms three primary germ layers, the mesoderm, the ectoderm, and the endoderm. These germ layers interact forming all the tissues and organs of the developing embryo. The influences controlling the transition of ectoderm to visceral and parietal endoderm in the blastocyst, followed by the formation of mesoderm at gastrulation, are only beginning to be defined. In the mouse, this process occurs between d 3 and 7 post-fertilization, and as such, it is both difficult to monitor and to experimentally influence. With this in mind, many groups have used mouse embryonic stem (ES) cells, and more recently human ES cells, to study the control of germ layer formation and their subsequent differentiation.
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