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

TY - JOUR

T1 - Optimization of three-dimensional imaging on in vitro produced porcine blastocysts and chimeras for stem cell testing

T2 - a technology report.

AU - Secher, Jan Ole Bertelsen

AU - Freude, Kristine

AU - Li, Rong

AU - Callesen, Henrik

PY - 2015/5/1

Y1 - 2015/5/1

N2 - 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.

AB - 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.

U2 - 10.1089/scd.2014.0503

DO - 10.1089/scd.2014.0503

M3 - Journal article

C2 - 25567670

SN - 1547-3287

VL - 24

SP - 1141

EP - 1145

JO - Stem Cells and Development

JF - Stem Cells and Development

IS - 9

ER -