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High-throughput screening method for progesterone response defect infertility human sperm samples

A screening method and high-throughput technology, applied in the medical field, can solve the problems of inefficient screening of sterile samples, cumbersome steps, and long time consumption, and achieve stable results, high sensitivity, and low photobleaching effects

Inactive Publication Date: 2020-03-10
NANTONG UNIVERSITY
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Problems solved by technology

[0005] Aiming at the cumbersome steps and long time-consuming of the existing methods, which fail to efficiently screen infertile samples with progesterone response defects, the present invention proposes a high-throughput screening method for infertile human sperm samples with progesterone response defects. Progesterone membrane receptors in sperm and the specific mechanism of action of progesterone on sperm and its clinical significance in evaluating the etiology of male infertility, and provide new ideas for finding effective ways to treat infertility

Method used

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  • High-throughput screening method for progesterone response defect infertility human sperm samples
  • High-throughput screening method for progesterone response defect infertility human sperm samples

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Embodiment 1

[0032] Step 1, Sperm Sample Processing

[0033] The semen samples collected on the day were liquefied in a constant temperature water bath at 37°C for 60 min. The liquefied semen samples were analyzed in the human semen analysis plate. Take 10 µL of the liquefied semen, place it on a clean 20 µm glass slide, and cover it with a cover glass to form a pool about 20 µm deep. Put the semen analysis plate The parameters of human sperm are detected in a computer-aided semen analyzer (CASA), the sperm samples are screened under a microscope, and fully liquefied sperm samples with good forward movement and normal morphology are selected as much as possible.

[0034] Step 2, upstream purification and isolation of human sperm

[0035] Human semen is isolated and purified by upstream methods. Take a 50 mL centrifuge tube, add 5 mL of the prepared HTF solution, and preheat at 37°C for 30 min for later use. Take out the preheated HTF, draw 1mL sperm sample with a pipette gun, insert the...

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Abstract

The invention discloses a high-throughput screening method for progesterone response defect infertility human sperm samples. According to the invention, healthy reproductive men are used as normal control, a male patient with infertility caused by unknown reasons is used as a research object, the effect of progesterone among different human sperm samples on the concentration ([Ca<2+>]i) of free Ca<2+> in head cells is observed and screened at high throughput, human sperm infertility samples with progesterone response defects are efficiently and rapidly screened while the dosing time differenceis avoided, the influence of progesterone on the human sperm function and the molecular mechanism of progesterone are further explored, and meanwhile, the clinical significance of sperm membrane progesterone receptor function defects in evaluation of male infertility causes is expected to be illuminated.

Description

technical field [0001] The invention belongs to the field of medical technology, and in particular relates to a high-throughput screening method for progesterone response-deficient infertile human sperm samples. Background technique [0002] After entering the female reproductive tract, sperm can quickly respond to changes in the external physical and chemical environment, and undergo a series of physiological processes and functional changes such as capacitation, hyperactivation, and acrosome reaction, before finally combining with egg cells to complete fertilization. Progesterone is the most important physiological hormone that regulates the function of mammalian mature sperm. It is secreted by cumulus cells and presents a concentration gradient distribution in the female reproductive tract. The physiological function acts as an inductive feature, guiding the sperm to successfully swim to the egg cell, and avoiding the premature hyperactivation of the sperm and the loss of...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64G01N33/58
CPCG01N21/6428G01N33/582
Inventor 曾旭辉顾亚云陈晨彭利忠杨会会张小宁
Owner NANTONG UNIVERSITY
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