Method of performing scRRBS analysis on embryo culture solution

Abstract

The invention provides a method of performing scRRBS (single-cell reduced-representation bisulfite sequencing) on an embryo culture solution. According to the method provided by the invention, medicalwaste (blastula culture solution) generated by 'test-tube baby' operation is adopted as a raw material, double analysis can be performed on the chromosome aneuploidy status and DNA methylation statusof the embryo, development potential of the embryo is evaluated from a brand new angle of epigenetics and reaction between the embryo and the culture environment, new reference is provided for selecting a 'correct' embryo in assisted reproduction, and powerful support is provided for increasing the success rate of the period of the test-tube baby.

Description

technical field [0001] The invention relates to the fields of biomedicine and molecular cell biology, in particular to a method for performing single-cell reduced-representation bifulfitesequencing (scRRBS) on embryonic culture fluid, which utilizes epigenetic The chemical analysis technology analyzes the embryo chromosome status and DNA methylation status in the blastocyst culture medium. Background technique [0002] Since July 1978, the first "test-tube baby" Louis Brown was born, and its founder Robert G. Edwards won the Nobel Prize in Medicine and Physiology in 2010. In the past forty years, millions of test-tube babies have been born around the world. The term "test-tube baby" has become widely known. Although with the rapid development of technology, the test tube baby technology has also gone through one generation and two generations, and now it has entered the third generation, but the overall success rate is still not up to people's expectations, and the current ...

AI Technical Summary

Problems solved by technology

In the early stage of embryonic development, the epigenetic status of the embryo undergoes drastic changes, which may be affected by the external environment and cause mutations, thereby affecting the developmental potential of the embryo, and leading to some embryos with high-quality morphology and normal chromosomal status. Epigenetic disturbances reduce developmental potential and lead to post-implantation failure

In the current preimplantation screening, there is no analysis of the epigenetic status of embryos.

[0006] At the same time, the current mainstream PGS uses trophoblast cell biopsy at the blastocyst stage. Although this biopsy has not been proven to be harmful to embryos and test-tube babies, it requires high technical requirements, and improper operation will cause consequences such as embryo suspension.

[0007] CN105368936A uses the waste liquid after embryo culture as a material, trying to achieve non-invasive detection, but still only analyzes the chromosome state

[0009] Biopsy sampling has high requirements on embryo operation techniques. Even if the operation is good, it will cause certain damage to the embryo. If the operation is wrong, it is likely to damage the embryo and cause serious consequences.

[0010] There may be chimerism in trophoblast cells, and a small number of 3-5 cells taken may not fully represent the embryo, resulting in misjudgment

[0011] Evaluation only from the chromosomal state ignores the very important epigenetic changes in the early embryo, and cannot provide more references for a more comprehensive evaluation of the developmental potential of the embryo

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