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Journal of Cell Science 115, 329-339 (2002)
© 2002 The Company of Biologists Limited


Research Article

Transient pluripotent cell populations during primitive ectoderm formation: correlation of in vivo and in vitro pluripotent cell development.

T. A. Pelton*, S. Sharma*,{ddagger}, T. C. Schulz*,§, J. Rathjen and P. D. Rathjen

Department of Molecular Biosciences, University of Adelaide, Adelaide 5005, South Australia
* Contributed equally to this work
{ddagger} Current address: Department of Ophthalmology, Flinders University of South Australia, Bedford Park, SA 5042, Australia
§ Current address: Laboratory of Cellular and Developmental Biology, Section of Mammalian Development, NIDDK, NIH, Bethesda, MD 50892, USA

¶Author for correspondence (e-mail: peter.rathjen{at}adelaide.edu.au)

Accepted September 24, 2001

Formation and differentiation of a pluripotent cell population is central to mammalian development, and the isolation, identification and manipulation of human pluripotent cells is predicted to be of therapeutic use. Within the early mammalian embryo, two distinct populations of pluripotent cells have been described: the inner cell mass (ICM), which differentiates to form a second pluripotent cell populations, the primitive ectoderm. Indirect evidence suggests the existence of temporally distinct intermediate pluripotent cell populations as primitive ectoderm is formed. We coupled an in vitro model of primitive ectoderm formation (the transition of embryonic stem cells to early primitive ectoderm-like (EPL) cells) with ddPCR-based techniques to identify three novel genes, Psc1, CRTR-1 and PRCE, that were expressed differently during pluripotent cell progression. Detailed mapping of these genes with Oct4, Rex1 and Fgf5 on pregastrulation embryos provided the first molecular evidence for the existence of successive, temporally distinct pluripotent cell populations in the embryo between the ICM and primitive ectoderm. No evidence was found for spatial heterogeneity within the Oct4+ pool. The transition between populations correlated with morphological or developmental alterations in pluripotent cells in vivo. Genes that are temporally expressed during pluripotent cell progression may provide an opportunity for molecular discrimination of pluripotent cells at different stages of maturation in vivo and an understanding of the cellular origins and properties of pluripotent cell lines isolated from diverse sources. Furthermore, the strong correlation of gene expression demonstrated between EPL cell formation in vitro and primitive ectoderm formation in vivo validates EPL cells as a model for primitive ectoderm, thereby providing a model system for the investigation of pluripotent differentiation and an opportunity for directed differentiation of pluripotent cells to therapeutically useful cell populations.

Key words: Pluripotent, ES cells, EPL cells, ICM, Primitive ectoderm, Mammalian development, Psc1, CRTR-1, PRCE, Differential gene expression


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