Sperm cryopreservation method based on protective agent

Abstract

The invention discloses a sperm cryopreservation method based on a protective agent, which comprises the following steps: (a) diluting a sperm sample by using a semen-rare sperm cryoprotectant, the volume ratio of the semen-rare sperm cryoprotectant to the sperm being 1: (1-2); (b) placing the semen-rare sperm cryoprotectant mixture at room temperature to act for at least 5 minutes; (c) putting the semen-rare sperm cryoprotectant mixture into a preservation container, buckling the container on a bracket, and then carrying out fumigating on a liquid nitrogen surface for 10 minutes; and (d) transferring the bracket loaded with the preservation container into a liquid nitrogen storage tank at -196 DEG C for storage. The liquid nitrogen storage tank in the step (d) comprises a tank body, a first vacuum layer, supporting legs, a freezing bin, a mounting opening, a freezing frame, a safety fastener, a lifting device, a clamping device and a two-axis coordinate positioning device. The freezing frame is arranged, so that more frozen test tubes can be stored in order. The two-axis coordinate positioning device is arranged, so that code storage and positioning rapid extraction can be realized, and the efficiency is improved.

Description

technical field [0001] The invention belongs to the technical field of assisted reproduction, and in particular relates to a method for cryopreserving sperm based on a protective agent. Background technique [0002] Semen freezing is an assisted reproductive technology, which is often used for artificial insemination in farmed animals or infertility assisted reproduction in humans; in the process of semen freezing, the artificially collected semen needs to be tested for virus resistance and sperm motility sampling After inspection, add a protective agent, seal it in a frozen test tube, and store it in an ultra-low temperature environment of liquid nitrogen; when necessary, take the semen out of the liquid nitrogen tank and thaw it, and then it can be used for fertilization; In the current liquid nitrogen freezing device, the structure of the liquid nitrogen device is relatively simple. Usually, a multi-layer drawer made of metal is hung on the mouth of the liquid nitrogen ta...

AI Technical Summary

Problems solved by technology

[0002] Semen freezing is an assisted reproductive technology, which is often used for artificial insemination in farmed animals or infertility assisted reproduction in humans; in the process of semen freezing, the artificially collected semen needs to be tested for virus resistance and sperm motility sampling After inspection, add a protective agent, seal it in a frozen test tube, and store it in an ultra-low temperature environment of liquid nitrogen; when necessary, take the semen out of the liquid nitrogen tank and thaw it, and then it can be used for fertilization; In the current liquid nitrogen freezing device, the structure of the liquid nitrogen device is relatively simple. Usually, a multi-layer drawer made of metal is hung on the mouth of the liquid nitrogen tank through a hook, and the metal drawer and the frozen test tubes stored therein are submerged in liquid nitrogen. , and then seal the liquid nitrogen tank through the gland; this liquid nitrogen freezing method has great disadvantages. First, because the hook is hung on the mouth of the tank, the gland has poor airtightness when closing the mouth of the liquid nitrogen tank. At the same time, due to the leakage of liquid nitrogen, the

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