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Application of growth differentiation factor 9 in prediction of embryonic development potential

A technology of growth and differentiation factors and embryonic development, applied in the field of assisted reproduction, can solve the problems of large subjective factors, large differences, and inability to quantify embryo quality, and achieve the effects of shortening the treatment cycle, increasing the live birth rate, and high clinical significance

Pending Publication Date: 2021-12-24
李竞宇
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the commonly used embryo selection technology is embryo morphology scoring, which mainly judges the quality of embryos by evaluating the development speed, blastomere size, fragments and vacuoles of embryos. The disadvantage of this method is that the subjective factors of evaluation are relatively large, and different techniques There are large differences between personnel and different laboratories, and the quality of embryos cannot be quantified
In addition, the morphology of the embryo sometimes does not fully reflect the implantation potential of the embryo

Method used

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  • Application of growth differentiation factor 9 in prediction of embryonic development potential
  • Application of growth differentiation factor 9 in prediction of embryonic development potential
  • Application of growth differentiation factor 9 in prediction of embryonic development potential

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] 1. Obtain the culture medium of the embryo: the embryo after fertilization is put into the pre-balanced cleavage stage embryo culture drop (G1+5% HSA recombinant human serum albumin) to continue to cultivate, the culture environment is 37 ℃, 5% oxygen and 6% carbon dioxide, embryo evaluation is based on traditional morphological scoring, including cleavage speed, fragmentation degree, blastomere uniformity, and blastomere appearance of mononucleation or multinucleation, etc., and the transferable embryos on the third day are defined as the number of blastomeres Embryos with no less than 6 fragments, less than 30% fragments and no obvious morphological abnormalities were obtained with good-quality embryo culture medium and poor-quality embryo culture medium.

[0023] 2. GDF9 (growth factor 9) content in the embryo culture fluid with good quality and bad quality embryo culture fluid of detection quality: determine the absorbance of described embryo culture fluid with good ...

Embodiment 2

[0030] Example 2. A kit for predicting or assisting in predicting embryo developmental potential

[0031] 1. The components of the kit: detection sample, standard substance, quality control substance and biotin, concentrated washing solution A, GDF-9 test buffer, biotin conjugate and TMB substrate solution.

[0032] 2. GDF9 detection method:

[0033] 1. Equilibrate all reagents and samples to room temperature;

[0034] 2. Use 1mL deionized water to dissolve standard products B-F (respectively containing recombinant human GDF9 protein 48pg, 222pg, 775pg, 2550pg, 5800pg) and quality control products I and II (respectively containing recombinant human GDF9 protein 240-400pg, 2868 -4302pg), dissolved and vortexed;

[0035] 3. Dilute the concentrated lotion A (phosphate buffer containing non-ionic detergent) 25 times with deionized water;

[0036] 4. Dilute the sample to an appropriate multiple with fresh blank culture medium (cleavage stage embryo culture medium);

[0037] 5. ...

Embodiment 3

[0052] Example 3. A method for predicting the developmental potential of embryos for non-therapeutic and diagnostic purposes

[0053] 1. select respectively the embryo culture fluid of several people who want to implant the same person, and the source and period of the embryo culture fluid are identical with embodiment 1;

[0054] 2. Use the kit of Example 2 to detect the content of growth factor-9 in the embryo culture medium in step 1, and select the embryo with the lowest content of growth factor-9 as the embryo with the most developmental potential.

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Abstract

The invention provides application of a growth differentiation factor 9 in prediction of embryonic development potential and belongs to the technical field of assisted reproduction. The invention aims to provide a safe and non-invasive marker of embryonic development potential and relates to a method for predicting the embryonic development potential by detecting the GDF9 level in a human embryo in-vitro culture solution. The GDF9 content in the embryo culture solution on the third day is obviously related to the embryonic development potential. The method comprises the following steps of: obtaining an embryo culture solution; determining the absorbance of the embryo culture solution; determining the absorbance of a culture solution without embryos; adopting the absorbance of a GDF9 standard substance solution and the concentration of the GDF9 standard substance as a standard curve; and comparing the absorbance of the embryo culture solution with the standard curve to obtain the concentration of the GDF9 of the embryo culture solution; and evaluating the embryo development potential by using the concentration of the GDF9. The biopsy of an embryo is not needed, so that the method not only has the advantage of noninvasiveness, but also can improve the living yield of a single assisted reproduction treatment cycle, and has relatively high clinical significance.

Description

technical field [0001] The invention belongs to the technical field of assisted reproduction, and in particular relates to the application of growth differentiation factor 9 in predicting the developmental potential of embryos. Background technique [0002] With the advancement of biological sciences, assisted reproductive technologies (ART) have made great progress in recent years, especially in IVF laboratory technology, but the clinical outcomes are still not satisfactory. According to the latest 2017 national ART data released by the US CDC, the implantation rate of transplanted embryos is only 37.1% for patients younger than 35 years old; as the age increases, the implantation rate of embryos also decreases, respectively: 27.5% (35 -37 years old), 18.3% (38-40 years old), 9.7% (41-42), 4.1% (43-44), among female assisted conception patients over 45 years old, embryos rarely implanted in 2.2%. [0003] Current research shows that embryo selection is an important factor ...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/569C12N5/073
CPCG01N33/74G01N33/56966C12N5/0604G01N2333/495C12N2501/998
Inventor 李竞宇
Owner 李竞宇
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