A method for cryopreservation of micro-sperm

Abstract

The invention discloses a microsperm cryopreservation method, aiming to provide a microsperm with small volume, closed space, easy operation and high sperm activity (including a single sperm, such as testicular puncture sperm, epididymal puncture sperm, severe The cryopreservation carrier and cryopreservation method of the sperm of patients with oligospermia and asthenospermia, etc.) When the sperm is recovered, take out the cryopreservation tube from the liquid nitrogen tank, take out the cryopreservation carrier from the cryopreservation tube with a straw, and quickly add it to the sperm culture dish Medium (37°C), under an inverted microscope (100-200 times magnification), carefully search for sperm in the frozen carrier, use the intrafollicular single sperm injection needle to grab the sperm and transfer them to the sperm culture medium of the sperm culture dish for subsequent sperm ICSI of immobilized and matured eggs.

Description

technical field [0001] The present invention relates to the field of assisted reproduction in medicine, in particular to cryopreservation techniques for trace sperm (single or dozens of sperm), including testicular puncture sperm, epididymal puncture sperm, and sperm from patients with severe oligospermia and asthenospermia. Background technique [0002] Infertility is a major disease that seriously endangers human reproductive health. Its incidence accounts for about 1 / 6 of couples of childbearing age in the world, and about 50% of them are caused by male factors. Sperm or azoospermia, wherein azoospermia accounts for about 15% of male infertility patients. There is no sperm in the semen of patients with azoospermia, including obstructive azoospermia (OA) and non-obstructive azoospermia (NOA). NOA patients are mainly caused by spermatogenesis or maturation disorders. Currently, there is no treatment to restore spermatogenesis in patients, accounting for about 60% of azoosp...

AI Technical Summary

Problems solved by technology

However, this method requires a special wet box under saturated humidity conditions to ensure that the droplets do not evaporate during the freezing process. The operation process is cumbersome and time-consuming, which greatly limits the wide application of this method in clinical practice.

In 2011, Lu Hui and Peng Qiuping developed a moisturizing device for freezing rare sperm. This moisturizing device provides constant humidity conditions. During the sperm transfer process, it can ensure that the humidity of the transfer environment remains unchanged and prevent sperm from being removed from the petri dish. In the process of transferring to a frozen carrier (such as a frozen sheet), the osmotic pressure in the liquid increases due to the evaporation of the droplets, causing sperm damage or even death, and reducing sperm activity. However, this device is only a moisturizing device and not a frozen carrier loaded with sperm. sperm cryopreservation

However, this method also has some problems. Firstly, when freezing and thawing, the liquid droplets are in direct contact with mineral oil and need to be cleaned and removed. If the removal is not clean, it may have an impact on sperm activity and ICSI treatment results. Secondly, the droplet volume is relatively large (0.1 -10.0μL), it takes a lot of time to find and grab the cryopreserved sperm when recovering the sperm by micromanipulation after thawing, thus limiting its clinical application

[0004] In summary, the current micro-sperm cryopreservation methods have certain defects, which limit their clinical application. Therefore, it is urgent to develop a sperm cryopreservation carrier with a smaller volume and a closed space.

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