Sperm freezing and unfreezing method and sperm freezing and unfreezing device

A technology of sperm and frozen slices, applied in biochemical equipment and methods, tissue cell/virus culture devices, microorganisms, etc., can solve the problems of low sperm count, sperm loss, low motility, etc., and achieve simple and easy operation, The effect of increased flexibility

Inactive Publication Date: 2013-05-29
SHANGHAI NINTH PEOPLES HOSPITAL SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, in the above method, frozen straws, empty zona pellucida, frozen rings, etc. are used to load sperm, and many sperm are lost during the operation, resulting in a small number of sperm obtained and low motility. After routine cryopreservation and recovery, it is often difficult to find Live sperm for ICSI

Method used

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  • Sperm freezing and unfreezing method and sperm freezing and unfreezing device
  • Sperm freezing and unfreezing method and sperm freezing and unfreezing device
  • Sperm freezing and unfreezing method and sperm freezing and unfreezing device

Examples

Experimental program
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Effect test

Embodiment 1

[0035] Take a small amount of normally discharged sperm and put it into a small amount of modified human fallopian tube fluid (mHTF), put the dish containing the sperm into the wet box 1, make a droplet of freezing liquid on the slice 7 of the frozen slice 6, and micromanipulate Use a needle to draw 20 sperm into the droplet. At this time, check the survival rate before freezing, and then put a pre-moistened freezing jacket 9 on the frozen sheet 6, slowly put the freezing jacket 9 into liquid nitrogen vapor, and expose it for 5 minutes Afterwards, they were directly put into liquid nitrogen for long-term cryopreservation.

[0036] When recovering, remove the freezing jacket 9 in liquid nitrogen, place the freezing piece 6 in mineral oil at 37 degrees Celsius, move slowly, cut off the piece 7 of the freezing piece 6 with scissors, and place a small weight (not shown) close to the freezing point. The liquid droplet presses the frozen sheet 6. Under the inverted microscope, use ...

Embodiment 2

[0038] Place the seminiferous tubules removed from the biopsy into a small amount of modified human fallopian tube fluid (mHTF), crush the seminiferous tubules with curved forceps, put the dish containing the seminiferous tubules into the wet box 1, Make a microdroplet of freezing liquid in the sheet part 8 of the frozen sheet 6, add mineral oil to cover and moisturize, use an injection needle to draw 10 spermatozoa and put them into the droplet, at this time, check the survival rate before freezing, and then put it on the frozen sheet 6 Put on the pre-wetted freezing jacket 9, slowly put the freezing jacket 9 into the liquid nitrogen vapor, and after exposing for 5 minutes, put it directly into the liquid nitrogen for long-term cryopreservation.

[0039] When recovering, remove the freezing jacket 9 in liquid nitrogen, place the frozen sheet 6 in mineral oil at 37 degrees Celsius, move slowly to rewarm, cut off the slice part 8 of the frozen sheet 6 with scissors, and place a ...

Embodiment 3

[0041] Place the seminiferous tubules removed from the biopsy into a small amount of modified human fallopian tube fluid (mHTF), crush the seminiferous tubules with curved forceps, put the dish containing the seminiferous tubules into the wet box 1, Make a droplet of freezing liquid in the sheet part 8 of the frozen sheet 6, add mineral oil to cover and moisturize, draw 20 spermatozoa into the droplet with an injection needle, at this time, check the survival rate before freezing, and then put it on the frozen sheet 6 Put on the pre-wetted freezing jacket 9, slowly put the freezing jacket 9 into the liquid nitrogen vapor, and after exposing for 5 minutes, put it directly into the liquid nitrogen for long-term cryopreservation.

[0042] When recovering, remove the freezing jacket 9 in liquid nitrogen, place the frozen sheet 6 in mineral oil at 37 degrees Celsius, move slowly to rewarm, cut off the slice part 8 of the frozen sheet 6 with scissors, and place a small weight (not sh...

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Abstract

The invention provides a sperm freezing and unfreezing method with simple and convenient operation, easy searching of sperms, high sperm survival rate and high sperm recovery rate, and also provides a device using the method. The method and the device have the main advantages that: (1) sperms are put into microdroplets and unfrozen in oil, the sperms cannot be lost, and the sperm recovery rate isapproximate to 100 percent; (2) refrigerating fluid contains glycerin, and the sperm survival rate can reach 90 percent; and (3) the method and the device have more simple, convenient and feasible operation than the prior art. The method and the device well solve the problem of easy failure of sperm recovery after severe oligo-asthenosperms, testicular sperms and epididymal sperms are frozen in the prior sperm freezing technology, and greatly improve the flexibility for azoospermia patients to receive assisted reproductive treatment.

Description

technical field [0001] The invention relates to the field of assisted reproduction in medicine, and relates to a method for freezing and thawing a small amount of testicular sperm and epididymal sperm and a device used in the method. Background technique [0002] Modern assisted reproductive technology can treat patients with oligospermia or azoospermia (a small amount of sperm obtained through biopsy of testis and epididymis) to obtain offspring by intracytoplasmic sperm injection (ICSI). Since the pregnancy rate of each ICSI treatment cycle is 35%-40%, in order to obtain pregnancy, some patients need to repeat ICSI treatment several times. If the sperm is not cryopreserved, ejaculation or testicular and epididymis biopsies are performed every cycle. The sperm of oligospermia is unstable, and the sperm may not be obtained, resulting in the failure of this cycle. However, biopsy of the testis and epididymis is a traumatic operation. Repeated operations not only bring additi...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/08C12M3/00
Inventor 匡延平彭秋平
Owner SHANGHAI NINTH PEOPLES HOSPITAL SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE
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