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Method for analyzing embryo quality based on molecular immunoassay of mammalian blastulas

A technology for mammalian and immune detection, applied in measuring devices, analytical materials, biological testing, etc., can solve problems such as inability to accurately predict cell differentiation fate, and achieve the effects of high accuracy, simple method, and simple operation

Inactive Publication Date: 2012-07-25
NORTHWEST A & F UNIV +1
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Problems solved by technology

The use of this method can better evaluate the quality of blastocysts, but there are also great defects. This method can only determine the location and cell number of the two, and cannot accurately predict the differentiation fate of cells at the molecular level.

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  • Method for analyzing embryo quality based on molecular immunoassay of mammalian blastulas

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Embodiment Construction

[0036] The invention provides an embryo quality analysis method based on molecular immunodetection of mammalian blastocysts, by staining two key marker proteins (OCT4 and CDX2) specifically expressed in the blastocyst inner cell mass and trophectoderm respectively, According to the expression of these two proteins in blastocysts, the quality of blastocysts can be evaluated at the molecular level. In the following, the present invention will be further described in detail in combination with the specific dyeing process and dyeing result, which is an explanation of the present invention rather than a limitation.

[0037] A method for assessing the quality of mammalian blastocysts, comprising the following steps:

[0038] Due to the need for immunostaining of mammalian blastocysts, the blastocysts are first pretreated as follows:

[0039] a. Fixing mammalian blastocysts: placing the obtained mammalian blastocysts in 4% paraformaldehyde / PBS and fixing them at room temperature for...

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Abstract

The invention discloses a method for analyzing embryo quality based on molecular immunoassay of mammalian blastulas. The expressions of key transcription factors OCT4 (octamer-binding transcription factor 4) and CDX2 (caudal-related homeodomain transcription 2) respectively expressing in inner cell masses and trophectoderm cells are used as the basis for analysis of the mammalian blastulas, and cellular level and molecular level analyses are carried out based on the immunostaining results of OCT4 and CDX2, so that the conventional blastula quality assessment method is covered, and the lineage differentiation with higher accuracy is proposed to conduct assessment; the blastulas with a proper ratio of the number of the inner cell masses to the number of the trophectoderm cells and complete first lineage differentiation are used as quality blastulas. Compared with the conventional blastula quality assessment method, the method disclosed by the invention not only can assess the blastula quality at a cellular level, but also can accurately assess the blastula quality and subsequent development potential at a molecular level, and the method is simple and high in accuracy and can provide technical support for embryonic implantation.

Description

technical field [0001] The invention belongs to the technical field of embryo quality assessment, and relates to an embryo quality analysis method based on molecular immunodetection of mammalian blastocysts. Background technique [0002] Embryo quality can be used to evaluate the viability of embryos, which is related to the potential of embryo development, and this development potential is directly related to the implantation of embryos and whether they can form normal individuals. Therefore, in in vitro fertilization technology and somatic cell nuclear transfer technology, it is an important prerequisite to ensure the efficiency of embryo transfer to accurately evaluate the quality of embryos to select good embryos. [0003] The evaluation of embryo quality is multi-level. The speed of embryo development to blastocyst, the shape of blastocyst, whether the inner cell mass of blastocyst is obvious, the total cell number of blastocyst and the ratio of inner cell mass cell nu...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/53
Inventor 彭辉张涌
Owner NORTHWEST A & F UNIV
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