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Establishment of primitive endoderm stem cell lines in mice [coculture]

ABSTRACT: The mammalian blastocyst consists of three distinct cell types: epiblast, trophoblast, and primitive endoderm (PrE). Although stem cell lines that retain functional properties of epiblasts or trophoblasts have been established, establishment of stem cell lines that fully retain developmental potential of PrE has long been awaited. Here we report derivation of primitive endoderm stem (PrES) cells, which are capable to give rise functional PrE derived tissues and support fetal development of PrE-depleted blastocysts in chimeras, in mice. ES (#1M), TS cell (#1F), PrES (#6-5), and XEN (#6-5) cells were dissociated to single cells by incubation with 0.05% trypsin-EDTA at 37 °C for 3 min. The cells were resuspended in DMEM (Thermo Fisher Scientific) with 10% FBS, 1 × glutamine-penicillin-streptomycin, 0.1mM β-ME. The cells were replated alone or in combinations of each at a concentration of 20 cells per microwell (1.3×103 cells/dish) on 60mm EZSPHERE® SP dishes (Iwaki) and were placed at 37 °C and 5% CO2 for 1 day or 3 days.

ORGANISM(S): Mus musculus

PROVIDER: GSE159182 | GEO | 2021/12/31

REPOSITORIES: GEO

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Establishment of primitive endoderm stem cell lines in mice [scRNA-Seq]

Project description:The mammalian blastocyst consists of three distinct cell types: epiblast, trophoblast, and primitive endoderm (PrE). Although stem cell lines that retain functional properties of epiblasts or trophoblasts have been established, establishment of stem cell lines that fully retain developmental potential of PrE has long been awaited. Here we report derivation of primitive endoderm stem (PrES) cells, which are capable to give rise functional PrE derived tissues and support fetal development of PrE-depleted blastocysts in chimeras, in mice.

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Establishment of mouse primitive endoderm stem cell lines [scRNA-Seq II]

Project description:The mammalian blastocyst consists of three distinct cell types: epiblast, trophoblast (TB), and primitive endoderm (PrE). Although embryonic stem cells (ESCs) and trophoblast stem cells (TSCs) retain the functional properties of Epi and TB, the currently available extraembryonic endoderm cells (XENC) do not fully recapitulate the developmental potential of PrE. Here we report derivation of primitive endoderm stem cells (PrESCs) in mice. PrESCs express both PrE and pluripotency marker genes like founder PrE. These cells are efficiently incorporated into PrE upon blastocyst injection, generate functionally competent PrE-derived tissues, and support fetal development of PrE-depleted blastocysts in chimeras. Establishment of PrESCs therefore represents a significant step-forward in elucidating the mechanisms for PrE specification and subsequent pre- and post-implantation development.

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Establishment of primitive endoderm stem cell lines in mice [bulk]

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BMP4 signalling directs primitive endoderm-derived XEN cells to an extraembryonic visceral endoderm identity

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