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Embryo secreting type endogenous polypeptide PDBCM and application thereof

A secretory, endogenous technology, applied in the field of assisted reproduction, which can solve unsatisfactory problems

Active Publication Date: 2017-09-01
NANJING MATERNITY & CHILD HEALTH CARE HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although in recent years, people have made some progress in improving embryo culture medium through the discovery of some secreted factors, but the effect of increasing the rate of blastocyst formation and improving the quality of blastocysts is not satisfactory.

Method used

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  • Embryo secreting type endogenous polypeptide PDBCM and application thereof
  • Embryo secreting type endogenous polypeptide PDBCM and application thereof
  • Embryo secreting type endogenous polypeptide PDBCM and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] 1.1 Screening of secreted endogenous polypeptides related to blastocyst formation in human blastocyst culture medium

[0029] In order to avoid mutual interference between embryos, we carried out single embryo culture on human cleavage-stage embryos, and collected embryo culture fluid (divided into blastocyst-forming group and non-forming blastocyst group) according to the outcome of blastocyst formation. Peptideomics analysis was performed on the culture medium of embryos and the culture medium of embryos without blastocysts. A total of 3228 peptides were identified by mass spectrometry, of which 201 were significantly different (T-test, Pfigure 1 ).

[0030] 1.2 Functional analysis of protein precursors of differential polypeptides

[0031] In view of the fact that the endogenous polypeptides of the cells are mainly derived from the degradation of protein precursors by the intracellular proteasome pathway, IPA software was used to analyze the functions of the protein...

Embodiment 2

[0032] Example 2 Effect of Polypeptide PDBCM on Preimplantation Embryo Development in Mouse

[0033] The HERC2-derived peptide, namely PDBCM, was synthesized by chemical synthesis. The amino acid sequence was shown in SEQ ID NO.1. Single embryo culture was performed on fertilized mouse eggs, and different concentrations (2 μg / ml, 20 μg / ml) were added to the embryo culture medium (KSOM). ml) of PDBCM polypeptide stimulation: take 6-8 weeks old ICR mice, inject PMSG, inject hCG 48h later, and take samples 16h later; obtain hyaluronidase-digested granulosa cells from oviduct ampulla under a microscope, and operate liquid in vitro Wash the naked eggs in M2, place them in the KSOM culture medium balanced in 5% CO2 at 37 degrees in advance, and develop to two cells. In the morning of the next day, the two cells were added to the KSOM medium supplemented with different concentrations of PDBCM, and continued to culture until blastocysts. Observing its effect on the blastocyst formati...

Embodiment 3

[0034] Example 3 Comparison of content of polypeptide PDBCM in high-quality blastocyst culture medium and non-high-quality blastocyst culture medium

[0035] With the help of the PDBCM polypeptide standard, we compared the content of PDBCM in the culture medium of high-quality blastocysts and non-high-quality blastocysts by mass spectrometry. The results are shown in Figure 4 , it was found that the content of PDBCM in high-quality blastocyst culture medium was significantly higher than that of non-high-quality blastocysts (blastocysts were scored using the Gamder blastocyst grading method, blastocysts above 3BB on the fifth day were high-quality blastocysts, and blastocysts below 3BB were high-quality blastocysts. Blastocysts are non-high-quality blastocysts).

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PUM

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Abstract

The invention discloses embryo secreting type endogenous polypeptide PDBCM and application thereof. The amino acid sequence of the embryo secreting type endogenous polypeptide PDBCM is as shown in SEQ ID NO.1. The polypeptide PDBCM can enhance BIRC6 expression, reduce apoptosis proteins Smac and reduce caspase-9 expression, thus promoting formation of blastula. The embryo secreting type endogenous polypeptide PDBCM can be applied to preparation of a medicine capable of promoting the formation of the blastula and / or improving the quality of the blastula or preparation of blastula culture liquid in an in vitro fertilization-embryo transplantation process.

Description

technical field [0001] The invention belongs to the field of assisted reproduction, and relates to an embryo secreted endogenous polypeptide PDBCM and its application. Background technique [0002] After more than 30 years of development and improvement, conventional in vitro fertilization-embryo transfer technology has technically solved long-term medical problems such as infertility. However, in the conventional treatment of in vitro fertilization-embryo transfer, in order to ensure a certain clinical pregnancy rate, the Transfer of 2-3 cleavage-stage embryos is a common practice. Therefore, in the past 30 years, the overall twin pregnancy rate of the population has increased by 50%-70%, and the triple pregnancy rate has increased by 400%. However, multiple pregnancy has seriously threatened the safety of mothers and babies. In recent years, maternal complications during pregnancy have increased by 3- 7 times, neonatal mortality and morbidity increased by 4-10 times, grea...

Claims

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Application Information

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IPC IPC(8): C07K7/08C07K14/47C12N15/12A61K38/10A61K38/17A61P31/04C12N5/073G01N33/68
CPCA61K38/00C07K7/08C07K14/4723C12N5/0603G01N33/68G01N2410/00
Inventor 凌秀凤赵纯张军强曹善仁时晓丹陈梦茜王佳怡
Owner NANJING MATERNITY & CHILD HEALTH CARE HOSPITAL
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