1. Laboratory of Molecular Biology, National Institute of Neurological Disorders and Stroke, National Institutes of Health, Bethesda, Maryland 20892, USA
2. Mammalian Development Laboratory, Department of Zoology, University of Oxford, South Parks Road, Oxford OX1 3PS, UK
3. Stem Cell Biology Section, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20892, USA
4. These authors contributed equally to this work.

Correspondence to: Paul J. Tesar^1,2,4Ronald D. G. McKay¹ Correspondence and requests for materials should be addressed to R.D.G.M. (Email: mckayr@ninds.nih.gov) and P.J.T. (Email: paultesar@ninds.nih.gov).

The application of human embryonic stem (ES) cells in medicine and biology has an inherent reliance on understanding the starting cell population. Human ES cells differ from mouse ES cells and the specific embryonic origin of both cell types is unclear. Previous work suggested that mouse ES cells could only be obtained from the embryo before implantation in the uterus^{1, 2, 3, 4, 5}. Here we show that cell lines can be derived from the epiblast, a tissue of the post-implantation embryo that generates the embryo proper. These cells, which we refer to as EpiSCs (post-implantation epiblast-derived stem cells), express transcription factors known to regulate pluripotency, maintain their genomic integrity, and robustly differentiate into the major somatic cell types as well as primordial germ cells. The EpiSC lines are distinct from mouse ES cells in their epigenetic state and the signals controlling their differentiation. Furthermore, EpiSC and human ES cells share patterns of gene expression and signalling responses that normally function in the epiblast. These results show that epiblast cells can be maintained as stable cell lines and interrogated to understand how pluripotent cells generate distinct fates during early development.

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