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Method for detecting quality of assisted reproductive technology by mouse embryo array

A technology of assisted reproductive technology and detection method, which is applied in the field of improved rat fetal test to detect the quality of assisted reproductive technology, can solve problems such as inability to identify culture conditions, and achieve the effects of improving in vitro growth and development and improving quality

Inactive Publication Date: 2015-02-25
南京优而生物科技发展有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This traditional standard cannot identify the impact of different culture conditions, culture medium types, equipment consumables, operating procedures and other factors on the quality of assisted reproductive technology

Method used

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  • Method for detecting quality of assisted reproductive technology by mouse embryo array
  • Method for detecting quality of assisted reproductive technology by mouse embryo array
  • Method for detecting quality of assisted reproductive technology by mouse embryo array

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046]The present invention cultures the fertilized eggs of mice in several kinds of culture fluids commonly used in clinical practice at present, and analyzes the formation rate of blastocyst stage embryos and the number of cells of blastocysts, the total number of cells of embryos of different growth and development stages, and the positive results of blastocyst stage embryos. The number of cells expressing embryonic stem cell biological indicator UTF1 and trophoblast cells expressing CDX2. By comparing with the embryos that grew and developed at the same period, it can be clearly found that the fertilized eggs of mice grow in different culture solutions to produce embryos of different quality.

[0047] Mouse fertilized eggs need to meet the following conditions for embryo culture:

[0048] 1) Take 10 microliters of culture medium micro-droplets and cover them with about 2 mm thick mineral oil;

[0049] 2) 37°C constant temperature, moisturizing, and 5% O 2 + 5% CO 2 + ...

Embodiment 2

[0067] To compare and analyze the effects of culture medium commonly used in the market on blastocyst production rate and total cell number.

[0068] The fertilized eggs were cultured in KSOM, KSOM+AA and sIVF medium until 108 hours after hCG, and the embryo quality test was performed.

[0069] Fertilized egg culture: the culture conditions are 37°C constant temperature, moisture, and 5% O 2 + 5 % CO 2 + 90%N 2 The three in vitro culture fluids are KSOM commonly used in the market, KSOM + AA (KSOM added to contain all essential and non-essential amino acids) and sIVF (Sydney IVF culture fluid, including cleavage fluid and blastocyst fluid). Fertilized eggs are cultured in KSOM and KSOM+AA for 84 hours, and in sIVF culture medium, they are cultured in different stages, in the first 48 hours in cleavage fluid and then transplanted into blastocyst fluid. The embryo culture medium is compared with the sIVF embryo culture medium produced by COOK Company (this culture medium ...

Embodiment 3

[0072] Compared with the total cell number of embryos at the same time in vivo, the influence of the culture medium commonly used in the market on the total cell number of embryos at different growth and development stages was analyzed.

[0073] Divide the fertilized mice into three groups on the day when the suppository was seen, and collect the fertilized eggs in the two groups and culture them in KSOM and sIVF culture medium. 2 + 5 % CO 2 + 90%N 2 of mixed gas. In vivo contemporaneous embryos were collected from another group of mice. All three groups were assumed to be contemporaneous embryos at the same time after hCG injection. 48 hours after the injection of hCG, every 12 hours by using the "one-step cell counting kit" and detecting the total cell number of each embryo under a fluorescent microscope, and comparing the total cell number of the embryo in the culture medium with the total cell number of the embryo in the same period in vivo Compare. It was conclud...

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Abstract

The invention discloses an improved method and a kit for detecting quality of an assisted reproductive technology by a mouse embryo array, and particularly relates to a method for detecting the quality of the assisted reproductive technology (ART) by utilizing cell number at each stage of mouse embryo development and UTF1 positive cell number expressed by blastocyst in the mouse embryo assay (MEA), belonging to the technical field of cytology and biology. At present, based on the internationally common standard MEA, the following two standards are increased: (1) total number of cells contained in embryo at each 12-hour embryonic development period; (2) number of UTF1 positive cells expressed by the blastocyst and number of CDX2 cells expressed by trophoblast cell as well as a proportion of the UTF1 positive cells and the CDX2 cells to total cells of the blastocyst. By comparing the two standards for in-vitro cultivated and developed embryo with the in-vivo embryo at the corresponding period, relatively sensitive and effective means are provided for quality control, including optimizing in-vitro culture conditions and optimizing embryo culture liquor, of the detected assisted reproductive technology, and reference standards are provided for improving the assisted reproductive technology.

Description

technical field [0001] The invention relates to the field of reproductive biomedicine, in particular to an improved mouse fetal test method for detecting the quality of assisted reproductive technology. [0002] Background technique [0003] At present, the most critical restrictive factor of pre-implantation embryos after in vitro fertilization is that humans have not found a non-invasive method to determine the quality of embryos, so as to ensure that the transferred embryos are of the highest quality. Therefore, there are obvious artificial uncertain factors that have affected the development of assisted reproductive technology. [0004] Due to the particularity of assisted reproductive technology, research on various conditions, including embryo culture medium, cannot be done directly through human embryos, but through mice and rabbits as models. Therefore, the Mouse embryo assay (MEA) is currently used as a routine method for testing the quality of assisted reproducti...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/06G01N33/569
Inventor 金星亮郑菊芬
Owner 南京优而生物科技发展有限公司
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