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Human oocyte cryoprotectant

A cryoprotectant and oocyte technology, applied in the direction of germ cells, animal cells, vertebrate cells, etc., can solve problems such as spindle abnormality, low embryonic development potential, egg damage, etc., to inhibit the formation of ice crystals and improve development The effect of improving potential and blastocyst rate

Active Publication Date: 2016-08-24
爱科(天津)生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, sucrose is routinely used as a non-permeable cryoprotectant in the egg freezing medium, but the oocytes are prone to form ice crystals during the freezing process, which will damage the eggs and affect the rearrangement of chromosomes, resulting in spindle abnormalities. Therefore, the fertilization rate of frozen-thawed eggs is not high, the cleavage rate is low, and the embryo development potential is low.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Human oocyte cryoprotectant of the present invention comprises ovum freezing liquid and ovum thawing liquid, and described ovum freezing liquid comprises balancing liquid and freezing liquid, and it is composed of:

[0035] Balance fluid (EM): 60% HTF 1024 (V / V) + 20% SSS (V / V) + 10% PROH (V / V) + 10% EG (V / V),

[0036] Coolant (VM): 40% HTF 1024 (V / V) + 20% SSS (V / V) + 20% PROH (V / V) + 20% EG (V / V) + 0.65mol / L trehalose;

[0037] The ovum thawing liquid comprises resuscitating liquid, diluent-1, diluent-2 and cleaning liquid, and it is composed of:

[0038] Resuscitation solution (TM): 80% HTF 1024 (V / V) + 20% SSS (V / V) + 1.0 mol / L trehalose,

[0039] Diluent-1 (DM-1): 80% HTF 1024 (V / V) + 20% SSS (V / V) + 0.5 mol / L trehalose,

[0040] Diluent-2 (DM-2): 80% HTF 1024 (V / V) + 20% SSS (V / V) + 0.2 mol / L trehalose,

[0041]Cleaning solution (WM): 80% HTF 1024 (V / V) + 20% SSS (V / V).

[0042] The oocyte freeze-thaw technique is implemented as follows:

[0043] Freezing pro...

Embodiment 2

[0050] Only the human serum substitute SSS was adjusted to HSA, and the others were the same as in Example 1. In this example, a total of 70 oocytes were collected, 70 were thawed, and 64 survived. The survival rate after freezing and thawing was 91.43%, and 60 oocytes were fertilized. 93.75%, 58 cleavages, 96.67% cleavage rate, 39.67% blastocyst rate, and 45% clinical pregnancy rate after embryo transfer.

Embodiment 3

[0052] Only the human serum substitute SSS was adjusted to SPS, and the others were the same as in Example 1. In this example, a total of 70 oocytes were collected, 70 were thawed, and 63 survived. The survival rate after freezing and thawing was 90.0%, and 58 oocytes were fertilized. 92.06%, 56 cleavages, 96.55% cleavage rate, 39.26% blastocyst rate, and 45% clinical pregnancy rate after embryo transfer.

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PUM

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Abstract

The invention discloses a human ovocyte cryoprotectant. The human ovocyte cryoprotectant contains an ovum refrigerating fluid and an ovum thawing fluid, wherein the ovum refrigerating fluid contains a balance fluid and a refrigerating fluid; the ovum thawing fluid contains a resuscitation fluid, a diluent-1, a diluent-2 and a cleaning solution. The human ovocyte cryoprotectant is developed by combined application of the propanediol and glycol permeable cryoprotectants, and taking the trehalose (trehalose, alpha-D-glucopyranosyl alpha-D-glucopyranoside, C12H22O11) as a non-permeating agent. The non-permeating cryoprotectant suitable for freezing the human ovocyte is developed so that the formation of the ice crystal in the cell can be inhibited, the function of the frozen ovum can be protected and the developmental potentiality of the frozen ovocyte can be greatly improved. The clinical result proves that the fertility rate, the cleavage rate and the high quality blastocyst rate of the frozen ovocyte can be greatly improved.

Description

technical field [0001] The invention relates to a cryoprotectant for human oocytes and belongs to the field of reproductive medicine engineering. Background technique [0002] In January 2006, the first test-tube baby in my country and the second in the world was born in Peking University Third Hospital. At present, China's ART (Assisted Reproductive Technology) derivative technology has been in the forefront of the world in terms of application scope and technical level. With the increase in the global infertility population, the delay of female reproductive age, and the increase of cancer patients, the demand for female fertility preservation is also increasing. Egg freezing technology has also been gradually applied clinically. Compared with frozen embryos, on the one hand, freezing eggs can solve many religious, legal and ethical issues. On the other hand, for women whose ovarian function is damaged or even lost due to reproductive system diseases, such as Egg freezing...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/075
Inventor 曹云霞章志国
Owner 爱科(天津)生物技术有限公司
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