Kit for detecting mammal blastula inner cell mass (ICM)/trophectoderm (TE) ratio and cell apoptosis

Abstract

The invention provides a kit for detecting mammal blastula inner cell mass (ICM) / trophectoderm (TE) ratio and cell apoptosis. The kit comprises a PBS solution containing 0.5% of BSA, a PBS solution containing 2% of paraformaldehyde, a PBS solution containing 0.5% of Triton X-100 and 0.05% of Tween20, a PBS solution containing 10% of goat serum and 0.05% of Tween20, a mouse anti-CDX2 primary antibody, a rabbit anti-caspase-3 primary antibody, an isothiocyanic acid green fluorescein-labeled goat anti-mouse IgG secondary antibody, an Alexa Fluor 594 red fluorescein-labeled goat anti-rabbit IgG secondary antibody, and a PBS solution containing 20 micromoles per liter of H33342. The kit is designed based on an immunofluorescence technology, respectively utilizes a trophoblast cell surface antigen CDX2 and important caspase-3 produced in cell apoptosis as target points, respectively utilizes the primary antibodies respectively aiming at CDX2 and caspase-3 and the secondary antibodies respectively carrying the fluorescein labels as reactants for an antigen-antibody reaction, utilizes the DNA dye to dye the DNAs in cells and realizes synchronous detection of blastula ICM / TE ratio and cell apoptosis in the same embryo. The kit has good repeatability and high efficiency.

Description

technical field [0001] The invention relates to the field of cell biology and immunofluorescence technology, in particular to a kit for detecting the ICM / TE ratio and cell apoptosis of mammalian blastocysts. Background technique [0002] With the continuous expansion and deepening of embryo biotechnology research, whether it is in vitro production of embryos, embryo micromanipulation or embryo transfer, while improving the number of embryos, the quality of embryos has become more and more research hotspots. Blastocyst cells are divided into inner cell mass (ICM) and trophectoderm (TE), the ratio of the two (ICM / TE) or the total number of inner cell mass cells to blastocyst cells (total cell number, TCN) ratio (ICM / TCN) can scientifically measure the quality of embryos. Differential staining is a cell counting method that clearly distinguishes between the two types of cells described above. However, the traditional differential staining method has disadvantages such as low ...

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Problems solved by technology

[0009] In addition, the existing embryo quality evaluation technology needs to use embryos for ICM / TE counting and apoptosis analysis respectively, thus doubling the number of embryos used for embryo quality evaluation and reducing the detection efficiency; and it cannot fully reflect the same blastocyst. Two aspects of quality are one-sided

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