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Molecular and cellular characterization of bovine extra-embryonic primary cultured cells: an in vitro model to study trophoblast, endoderm and mesoderm crosstalks before implantation
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National audience. In mammals, extra-embryonic tissues are essential to support embryo patterning but also embryo survival, especially in late implanting species. These tissues are composed of 3 cell types: trophoblast, endoderm and mesoderm, each one being affected by somatic cell nuclear transfer (SCNT). At present it is unclear how these cells interact. In this study, we have established primary cell cultures of extra-embryonic tissues from bovine embryos collected at day-18 after artificial insemination. After trypsic digestions and percoll gradients, these extra-embryonic cells were cultured for up to a week on a plastic dish (endoderm, mesoderm) or matrigel/collagenIV coated dish (trophoblast). We validated each culture by comparing it to the corresponding cells from in vivo micro-dissected embryos (microarrays, immunofluorescence). These primary cell cultures were a powerful tool to start studying their cellular properties (CYTOOTM chip) and will further allow in vitro studies on cellular interactions among extra-embryonic tissues (among control cells or control vs. SCNT derived cells), and potentially between extra-embryonic vs. embryonic tissues.
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