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Culture medium and method for evaluating embryo quality by using culture medium

A technology of embryo quality and culture medium, applied in the field of protein detection, to achieve an effective and accurate evaluation method

Active Publication Date: 2020-01-14
深圳中山泌尿外科医院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

And this response may be closely related to the quality and implantation potential of BCM corresponding embryos
However, there is a lack of a method for evaluating the quality of embryos using BCM in the prior art.

Method used

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  • Culture medium and method for evaluating embryo quality by using culture medium

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] A culture medium, comprising fetal bovine serum albumin, insulin-transferrin-selenium additive and puromycin, the mass ratio of said fetal bovine serum albumin, insulin-transferrin-selenium additive and puromycin is 1979:20 :1.

[0031] Optionally, the concentration of the fetal bovine serum albumin is 10% g / dl, the concentration of the insulin-transferrin-selenium additive is 1% g / dl, and the concentration of the puromycin is 500 ng / mL.

[0032] A method for evaluating the quality of embryos using the above-mentioned culture medium, comprising the following steps:

[0033] A. Embryo culture and collection of embryo conditional culture medium: take 30 μ L of blastocyst culture medium droplets of high-quality blastocysts, take 10 g of fertilized eggs and use sequential culture medium to carry out embryo culture to obtain embryos, and the embryos are obtained according to the following procedures after 2 days of embryo culture. The blastocyst culture medium was added to ...

Embodiment 2

[0039] The difference from Example 1 is that in step A, 30 μL micro-droplets of blastocyst culture medium without blastocyst formation were taken.

Embodiment 3

[0041] The difference from Example 1 is that in step A, 30 μL of blastocyst culture fluid that has not been cultured before blastocysts is taken.

[0042] Comprehensive embodiment 1 to embodiment 3, obtained as figure 1 Differential protein assay plots of proteins secreted by endometrial cells are shown. The IL-6 content in the blastocyst culture medium droplet of high-quality blastocyst was detected by protein chip, and the fluorescence signal value was 1843.0; while the fluorescence signal value of IL-6 detected by the blastocyst culture medium droplet without blastocyst formation was 1080.5; The fluorescent signal value of IL-6 detected by micro-droplets of blastocyst culture fluid that had not been cultured was 478.4. This indicates that BCM with different blastocyst quality can induce endometrial cells to secrete IL-6 content as the basis for embryo quality assessment, and become a new non-invasive embryo quality assessment method.

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Abstract

The present invention belongs to the field of protein detection and particularly relates to a culture medium and a method for evaluating embryo quality by using the culture medium. The culture mediumcomprises fetal bovine serum proteins, an insulin-transferrin-selenium additive and puromycin. The method for evaluating the embryo quality by using the culture medium successively comprises the following steps of embryo culturing and embryo conditioned culture solution collecting, embryo conditioned culture solution thawing, endometrial stromal cell culturing, embryo conditioned culture solutionadding and protein detecting, differential protein of specific cell secreted protein is obtained and the embryo quality is evaluated. The evaluating method is very effective and accurate and makes upfor gaps in the market.

Description

technical field [0001] The invention belongs to the field of protein detection, and in particular relates to a culture medium and a method for evaluating embryo quality by using the culture medium. Background technique [0002] Embryo conditioned medium, referred to as BCM, is a culture medium containing embryos. In vitro fertilization-embryo transfer (invitrofertilizationandembryotransfer, IVF-ET) is currently the best way to treat infertility. The existing assessment method of embryos mainly scores embryos based on morphological parameters such as cell number, fragmentation and uniformity. However, in the actual operation process, it is found that this method is greatly affected by human subjective factors when grading embryos, and there are often inconsistent scores. The phenomenon. In order to further increase the success rate and reduce the multiple pregnancy rate, a more scientific method is needed to identify the quality of embryos, so as to reduce the number and fr...

Claims

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Application Information

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IPC IPC(8): C12N5/073G01N33/68
CPCC12N5/0604C12N2500/25C12N2501/998G01N33/68
Inventor 曾勇姚志鸿熊风孙青李观贵陈培林钟惠娴万才云彭晓琪周灿权
Owner 深圳中山泌尿外科医院
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