Trace sperm cryopreservation carrier and corresponding freezing and thawing methods thereof

A cryopreservation and carrier technology, which is applied in the preservation, application of human or animal body, and obstetrics and gynecology instruments, etc., can solve the problems of cumbersome operation process, difficult material source, and difficulty in being widely used in assisted reproduction clinics, and achieves freezing and thawing. Simple operation, increased flexibility and reliability

Inactive Publication Date: 2015-08-12
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

However, this method requires the zona pellucida of animal (mouse or rat) eggs or human eggs, and the source of materials is difficult (the zona pellucida of human eggs is difficult to obtain), and there are ethical disputes (the host DNA is injected into human eggs together with sperm during ICSI. risk), so it is difficult for this method to be widely used in assisted reproductive clinics
[0005] Our research group once invented a method for freezing and thawing micro-sperm and a device for freezing and thawing sperm (Chinese patent application 200810042695.4). , the sperm are frozen in the microdroplets on the frozen sheet, thawed in hot oil, and the sperm recovery rate is close to 100%, but this method needs to manufacture a special wet box with saturated humidity conditions to ensure that the microdroplets do not evaporate during the freezing process. It is cumbersome and time-consuming, which greatly limits the widespread clinical use of the invention

Method used

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  • Trace sperm cryopreservation carrier and corresponding freezing and thawing methods thereof
  • Trace sperm cryopreservation carrier and corresponding freezing and thawing methods thereof
  • Trace sperm cryopreservation carrier and corresponding freezing and thawing methods thereof

Examples

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Embodiment 1

[0024] Example 1. Microsperm cryopreservation carrier

[0025] See figure 1 , figure 1 As a preferred embodiment of the present invention, the cryopreservation carrier (also known as a cryopreservation sheet) is rectangular in size and can carry 1-3 freezing liquid droplets of 0.1-10.0uL, made of a transparent plastic material (such as polystyrene or Polypropylene) or transparent glass material, with a raised clamping end on one edge.

[0026] As a preferred solution, the cryopreservation carrier is rectangular, with a length of 0.3-1.5 cm and a width of 0.2-1.0 cm.

[0027] The shape of the cryopreservation carrier can be rectangular, square or circular, horizontal or groove-shaped, as long as it is convenient for long-term storage in a liquid nitrogen tank, the cryopreservation carrier can be stored in a container of any shape or on a support material for subsequent freezing procedures, such as placing the cryopreservation carrier into a 0.5-2.0mL cryovial or attaching th...

Embodiment 2

[0029] Example 2. The freezing and thawing method of microsperm

[0030] For freezing, prepare the sperm culture dish first by making long droplets with 10% SSS-mHTF and covering with mineral oil. The seminiferous tubules removed from the biopsy were placed in a small amount of 10% SSS-mHTF, the seminiferous tubules were crushed with curved forceps, and directly added to the long 10% SSS-mHTF droplet in the sperm culture dish. Prepare a 5-microliter droplet of freezing solution on the cryopreservation carrier, insert the cryopreservation carrier into a sperm culture dish covered with oil, and use a microinjection needle (ICSI Injection Pipette) to grab a certain number (5-10 spermatozoa) / piece) of motile sperm into the cryopreservation carrier micro-droplet, take out the cryopreservation carrier with tweezers, blot the mineral oil on the cryopreservation carrier with absorbent paper as much as possible, and put it into a 1.8mL cryovial (Vial) , in the order of 4°C for 20 mi...

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Abstract

The invention discloses a trace sperm cryopreservation carrier and corresponding freezing and thawing methods thereof. The trace sperm cryopreservation carrier is in a rectangular, square or circular horizontal or groove shape and is made of a transparent plastic or transparent glass, and the size of the trace sperm cryopreservation carrier can bear 1-3 0.1-10.0 mu L freezing liquid micro drops. The freezing and thawing method comprises the following steps: after preparing the freezing liquid micro drops on the trace sperm cryopreservation carrier, inserting the trace sperm cryopreservation carrier into a sperm culture dish covered with oil, grabbing sperms by a microinjection needle, putting into the freezing liquid micro drops, taking out the cryopreservation carrier, soaking up mineral oil by virtue of an absorbent paper, rapidly freezing, directly inserting the cryopreservation carrier into an ICSI vessel when thawing, and thawing the sperms by utilizing the mineral oil with the temperature of 37 DEG C. According to the trace sperm cryopreservation carrier and the corresponding freezing and thawing method thereof disclosed by the invention, as the cryopreservation carrier is inserted into the sperm culture dish covered with oil, and the sperms are grabbed and frozen, a special moisture retention device is not needed, and evaporation of the freezing liquid micro drops is not needed to be worried about; the operation is very simple, the flexibility and reliability of assisted reproductive treatment for a patient with azoospermia are greatly improved, the sperm recovery rate can reach 100 percent, and the sperm viability is about 60 percent.

Description

technical field [0001] The present invention relates to the field of assisted reproduction in medicine, more specifically, to a cryopreservation method and a cryopreservation carrier for trace sperm (including a single sperm, such as testicular puncture sperm, epididymal puncture sperm, sperm from patients with severe oligospermia, etc.) . Background technique [0002] In the clinic of assisted reproduction, patients with azoospermia usually undergo a diagnostic puncture of the epididymis or testis to ensure that the ovulation induction cycle can obtain enough sperm for intracytoplasmic sperm injection (Intracytoplasmic sperm injection, ICSI) treatment. Since the pregnancy rate of each cycle of ICSI treatment is only about 50%, in order to achieve pregnancy, some patients need to repeat ICSI treatment several times. In order to minimize the pain of repeated punctures and better preserve the spermatogenic function of men, medical workers try to cryopreserve the micromotile s...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N1/02A61B17/43
Inventor 薛松果彭秋平曹少锋匡延平
Owner SHANGHAI NINTH PEOPLES HOSPITAL SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE
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