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Method for evaluating or assisting in evaluating ovarian cells and marker used by method

An oocyte and auxiliary identification technology, applied in the field of biomedicine, can solve the problems of age-related decline in female reproductive capacity and no obvious improvement in the aging process of ovarian tissue.

Active Publication Date: 2020-03-31
INST OF ZOOLOGY CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although with the improvement and development of the medical level, the life span of human beings is gradually extending, but the aging decline of human female reproductive ability and the aging process of ovarian tissue have not been significantly improved

Method used

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  • Method for evaluating or assisting in evaluating ovarian cells and marker used by method
  • Method for evaluating or assisting in evaluating ovarian cells and marker used by method
  • Method for evaluating or assisting in evaluating ovarian cells and marker used by method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0111] Example 1. Acquisition of cynomolgus monkey single-cell sequencing library (including cynomolgus monkey oocyte sequencing library and cynomolgus monkey ovary somatic cell sequencing library)

[0112] Schematic diagram of the construction and analysis of cynomolgus monkey single-cell sequencing library, see figure 1 .

[0113] 1. Obtaining Cynomolgus Monkey Cell Suspension

[0114] According to the average life expectancy of cynomolgus monkeys and its corresponding relationship with human age, the young group consists of 4 (female) cynomolgus monkeys aged 4-5, and the old group consists of 4 (female) cynomolgus monkeys aged 18-20. Group. All cynomolgus monkeys underwent physical examination, including the measurement of basic parameters (such as body mass index (BMI)), the detection of biochemical indicators (such as blood routine, urine routine), and the examination of important internal organs such as brain MRI and heart. Ultrasonography was performed to confirm tha...

Embodiment 2

[0141] Example 2. Acquisition and detection of oocyte molecular markers

[0142] 1. Bioinformatics analysis to obtain oocyte molecular markers

[0143] Using the cynomolgus monkey genome Macaca_fascicularis_5.0 (Ensemble) as a reference, compare the sequencing data after quality control, count the unique comparison read length, and split it into different cell data. The cells are oocytes and somatic cells. Somatic cells Including stromal cells, smooth muscle cells, granulosa cells, endothelial cells, natural killer T cells, macrophages, etc.). A single molecule is marked with a unique molecular number, the amplification bias is corrected, and the single-cell expression map of cynomolgus monkey ovary is obtained. Search for highly variable genes through the Seurat software package, perform nonlinear dimensionality reduction (t-SNE) on these genes in the measured cells based on the gene group, perform clustering through the Find Clusters function of the Seurat software package ...

Embodiment 3

[0153] Example 3. Acquisition and detection of early oocyte molecular markers

[0154] Early oocytes are compact oocytes in primordial follicles surrounded by a single layer of flat granulosa cells, or larger oocytes in primary follicles surrounded by a single layer of cuboidal granulosa cells (secondary follicles). Middle and late oocytes are larger oocytes surrounded by at least a partial or complete second layer of cuboidal granulosa cells, or larger oocytes in cystic follicles with follicular fluid or cystic structures ( Antral follicles).

[0155] 1. Bioinformatics analysis to obtain early oocyte molecular markers

[0156] 1. PCA analysis of data from cynomolgus monkey oocyte sequencing library.

[0157] PCA analysis results see Figure 4 Middle A. The results showed that oocytes were divided into 4 subpopulations, C1-C4.

[0158] 2. Detect the relative expression of ZP1 gene, WEE2 gene, DNMT1 gene, ATP6 gene, FIGLA gene and SOX17 gene in the four subgroups respectiv...

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Abstract

The invention discloses a method for evaluating or assisting in evaluating ovarian cells and a marker used by the method. The experiment proves that at least one of LMOD3, RBM46 and NETO1 is used as the marker and can be used for identifying or assisting in identifying oocytes; REXO5 serving as the marker can be used for identifying or assisting in identifying early oocytes; GPX1 and / or GSR can beused as the markers for distinguishing the senescent oocytes and young oocytes, and at least one of IDH1, PRDX4 and NDUFB10 can be used as the markers for distinguishing senescent granulocytes and young granulocytes. The marker can be used for evaluating ovarian cancer, premature ovarian failure and other ovarian aging related diseases, and can also be used for detecting ovarian functions, oocyteviability and the like in a human assisted reproductive technology. The method has important application value.

Description

technical field [0001] The invention belongs to the field of biomedicine, and in particular relates to a method for evaluating or assisting in evaluating ovarian cells and a marker used therefor. Background technique [0002] As one of the most important organs in women, the ovary mainly functions in the mature discharge of oocytes, which depends on the signal interaction between the oocytes in the follicle and the surrounding granulosa cells and theca cells. Abnormal oocytes or surrounding somatic cells (including granulosa cells) will affect the normal function of the ovary. The ovary is an organ that undergoes degenerative changes relatively early in the aging process of the body. Ovarian aging and its accompanying reduction in female reproductive levels seriously affect the country's birth rate. Although with the improvement and development of the medical level, the life span of human beings is gradually prolonging, but the aging decline of human female reproductive abi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6888
CPCC12Q1/6888C12Q2600/158
Inventor 曲静刘光慧汤富酬王思张维绮李静宜郑宇轩于洋
Owner INST OF ZOOLOGY CHINESE ACAD OF SCI