Application of SLC2A1 expression to preparing kit for detecting fertility of oocytes

A technology for detecting kits and oocytes is applied in the field of evaluating the quality of oocytes and their fertilization ability, which can solve problems such as unclear regulation mechanism, and achieve the effect of simple and easy quality control means.

Active Publication Date: 2019-05-31
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

However, the regulatory mechanism of glucose metabolism in the process of oocyte development, fertilization and preimplantation embryo development is still unclear, especially the glucose transporters involved in glucose metabolism need to be further studied

Method used

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  • Application of SLC2A1 expression to preparing kit for detecting fertility of oocytes
  • Application of SLC2A1 expression to preparing kit for detecting fertility of oocytes
  • Application of SLC2A1 expression to preparing kit for detecting fertility of oocytes

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Experimental program
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Effect test

Embodiment 1

[0040] The present invention analyzes the situation of all known 12 glucose transporters (SLC2A) in mouse MII oocytes. from figure 1 In addition to SLC2A2, other 11 kinds of SLC2A were expressed. SLC2A1 is the most expressed glucose transporter, which is comparable to the expression of the housekeeping gene ACTB, while the others are less expressed.

[0041] The specific testing process includes:

[0042] 1) Choose female mice aged 6-8 weeks, and inject PMSG intraperitoneally, 10IU / mouse. After 48 hours, intraperitoneal injection of hCG, 10IU / cattle; 20 hours after HCG injection;

[0043] 2) Place the flocculent oocyte mass collected in the ampulla of the oviduct into hyaluronidase preheated at 37°C, and transfer immediately after the cumulus and granulosa cells around the embryo are digested and separated;

[0044] 3) Collect under a microscope. Rinse in pre-cooled PBS and repeat 3 times. 30 oocytes were transferred to 5 microliters of RNA buffer; all RNA was collected ...

Embodiment 2

[0056] Example 2 Comparative analysis of expression of SLC2A1 gene in mouse and human oocyte maturation process

[0057] from figure 2 found that SLC2A1 mRNA was already detected in GV stage oocytes, and SLC2A1 expression increased with maturation (p0.05).

[0058] The specific testing process includes:

[0059] 1) Choose female mice aged 6-8 weeks, and inject PMSG 10IU / rat intraperitoneally. After 48 hours, intraperitoneal injection of hCG 10IU / rat for superovulation, and 16-20 hours after HCG injection, they were killed;

[0060] 2) The flocculent oocyte clusters collected in the ampulla of the oviduct are placed in hyaluronidase preheated at 37°C, and transferred immediately after the cumulus and granulosa cells around the embryo are digested and separated;

[0061] 3) Human oocytes are derived from discarded oocytes voluntarily donated by assisted reproductive centers;

[0062] 4) Collect under a microscope. Rinse in pre-cooled PBS and repeat 3 times. Transfer a sin...

Embodiment 3

[0074] Relationship between fertilization ability and SLC2A1 in mouse MII oocytes. Fertilization rate and oocyte positive SLC2A1 expression rate ( Figure 3-1 ); comparison of SCL2A1 expression between fertilized eggs and unfertilized eggs ( Figure 3-2 ).

[0075] The specific testing process includes:

[0076] 1) Choose female mice aged 6-8 weeks, intraperitoneally inject PMSG 10IU / mouse; 48 hours later, intraperitoneally inject hCG 10IU / mouse for superovulation, inject HCG and share the cage overnight with male mice with proven fertility, 18 to 22 The female rats were put to death after 1 hour;

[0077] 2) The flocculent oocyte clusters collected in the ampulla of the oviduct were placed in hyaluronidase preheated at 37°C, and transferred immediately after the cumulus and granulosa cells around the embryo were digested and separated;

[0078] 3) Check the fertilized oocytes and unfertilized oocytes under a microscope, collect them separately; calculate the fertilization...

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Abstract

The invention discloses application of SLC2A1 expression to preparing a kit for detecting the fertility of oocytes, and a method for evaluating the physiological function of the oocytes, and belongs to the technical field of the cytology and biology. The application is uppermost characterized in that SLC2A1 is glucose transport protein the most expressed in the glucose transport protein family inthe oocytes, the SLC2A1 expression amount is in positive correlation with the fertility in the oocytes, the unfertilized oocytes present the missing and remarkable dropping of SLC2A1 genes, the kit for detecting the fertility of the oocytes detects and analyzes the expression of all the currently-known members in the same SLC2A family in the oocytes through the real-time fluorescent quantitative reverse transcription polypeptide chain reaction (qRT-PCR) method, it is found that the SLC2A1 is related to the quality and fertility of the oocytes, and the effective biological evaluation means is provided for controlling the quality of the assisted reproductive technology.

Description

technical field [0001] The invention relates to the fields of reproductive biology and reproductive medicine, in particular to a method for quantitatively detecting the expression of glucose transporter SLC2A1 in oocytes to prepare oocyte fertilization ability detection kits for evaluating the quality of oocytes and their fertilization A new approach to capacity. Background technique [0002] Glucose is an important energy source for mammalian cells. Glucose metabolism directly affects the reproductive ability of germ cells, and affects the embryonic development and implantation ability of blastocysts from fertilization to blastocysts (preimplantation embryos, PreimplantationEmbryo). Oocyte maturation includes nuclear maturation and cytoplasmic maturation, energy metabolism is crucial to its maturation, and oocytes obtain energy through various substrates, including carbohydrates, amino acids and fats. Studies have shown that glucose is an essential substrate for oocyte ma...

Claims

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Application Information

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IPC IPC(8): C12Q1/6876C12Q1/6851
Inventor 金星亮季煦韩倩倩常露杨五宁杨连增
Owner 南京优而生物科技发展有限公司
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