Optimization of three-dimensional imaging on in vitro produced porcine blastocysts and chimeras for stem cell testing: a technology report.

Jan Ole Bertelsen Secher, Kristine Freude, Rong Li, Henrik Callesen

    5 Citations (Scopus)

    Abstract

    Differential staining is an immunocytochemical staining that visualizes trophectoderm (TE) and the inner cell mass (ICM) of the blastocysts. It is used to determine the blastocyst quality, but could also be a useful tool to assess the integration site of injected cells into the early embryo. This is relevant for testing of presumed pluripotent stem cells. The gold standard for pluripotent stem cells is to test if the cells are capable of contributing to germline chimeras. Differential staining can be used to evaluate the possibility of chimeric contribution; if the cells are located in the area of the ICM they are likely to contribute to the fetus and if they are located in the area of the TE they are likely to contribute to the fetal membranes. In this article, we optimize on methods for embryo staining and mounting so that the exact location of injected stem cells within preimplantation porcine embryos can be evaluated.
    Original languageEnglish
    JournalStem Cells and Development
    Volume24
    Issue number9
    Pages (from-to)1141-1145
    Number of pages5
    ISSN1547-3287
    DOIs
    Publication statusPublished - 1 May 2015

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