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Application of PVA-based cryoprotectant in oocyte or embryo cryopreservation

A technology of oocyte and cryopreservation solution, which is applied in the application field of cryopreservation solution in oocyte or embryo cryopreservation, and can solve the problems that affect the safety and function expression of the preserved object, do not have ice crystal growth, damage cells, etc.

Active Publication Date: 2020-10-20
PEKING UNIV THIRD HOSPITAL +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, existing cryopreservation reagents cannot effectively control the growth of ice crystals during rewarming, which can damage cells
The high-concentration (≥15%) organic solvents used in the current vitrification method lead to toxic side effects of cryopreservation reagents on cells, seriously affecting the safety and functional expression of preserved objects (offspring)
In summary, the cryopreservation reagents currently used do not have the ability to effectively control the growth of ice crystals during the rewarming process, and there is also the problem of high reagent toxicity.

Method used

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  • Application of PVA-based cryoprotectant in oocyte or embryo cryopreservation
  • Application of PVA-based cryoprotectant in oocyte or embryo cryopreservation
  • Application of PVA-based cryoprotectant in oocyte or embryo cryopreservation

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preparation example Construction

[0100] The preparation method of the present invention will be further described in detail in conjunction with specific examples below. It should be understood that the following examples are only for illustrating and explaining the present invention, and should not be construed as limiting the protection scope of the present invention. All technologies realized based on the above contents of the present invention are covered within the scope of protection intended by the present invention.

[0101] Unless otherwise specified, the experimental methods used in the following examples are conventional methods; the reagents and materials used in the following examples, unless otherwise specified, can be obtained from commercial sources.

[0102] The PVA used in the embodiment of the present invention has a syndiotacticity of 50%-55%, a molecular weight of 13-23kDa, and a degree of hydrolysis of 98%.

[0103] In the cryopreservation solution in the embodiment of the present invent...

Embodiment

[0106] 1. Preparation of cryopreservation solution: prepare cryopreservation solution according to the following formula

[0107] Cryopreservation solution A:

[0108] Each 100mL contains the following components:

[0109] substance content PVA (g) 2.0 Ethylene glycol (mL) 10 DMSO (mL) 10 Sucrose (mol L -1 )

0.5 Fetal bovine serum (mL) 20 DPBS (mL) margin

[0110] 2.0g of PVA was heated in a water bath at 80°C and dissolved in 25mL of DPBS with magnetic stirring. After the PVA was completely dissolved and cooled to room temperature, the pH was adjusted to 7.0, which was solution 1; 17g (0.05mol) of sucrose (sucrose was frozen The final concentration in the preservation solution is 0.5mol L -1 ) was ultrasonically dissolved in 25mL of DPBS. After the sucrose was completely dissolved, 10mL of ethylene glycol and 10mL of DMSO were added to form solution 2. After solution 1 and solution 2 returned to room temperature, the t...

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Abstract

The invention discloses an application of a PVA-based cryoprotectant in oocyte or embryo cryopreservation. The cryoprotectant contains PVA, polyol and water-soluble sugar. PVA is used as a main cryoprotectant component of the cryoprotectant, the DMSO content is low, the toxicity is low, when the cryoprotectant is used for cryopreservation of oocytes, cell survival rate and functional expression stability are equal to or even higher than those of commercial cryoprotectant (containing 15% of DMSO), and the preservation efficiency is high. The cryoprotectant is simple in composition, the raw materials are easily available, and the cost is low.

Description

technical field [0001] The invention belongs to the technical field of biomedical materials, and in particular relates to the application of a PVA-based cryopreservation solution in oocyte or embryo cryopreservation. Background technique [0002] Cryopreservation (cryopreservation) refers to the preservation of cells, tissues or organs at ultra-low temperature, which slows down or stops the metabolism and division of cells, and can continue to develop once the normal physiological temperature is restored. Since the technology came out, it has become one of the indispensable research methods in the field of natural science and has been widely used. In recent years, with the increase of life pressure, human fertility has shown a downward trend year by year, and more and more people pay attention to fertility preservation. The cryopreservation of human germ cells (sperm, oocyte), gonad tissue, etc. important means of fertility. In addition, as the world's population ages, the...

Claims

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Application Information

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IPC IPC(8): A01N1/02
CPCA01N1/0221A01N1/0284
Inventor 乔杰严杰李蓉闫丽盈王健君金晟琳
Owner PEKING UNIV THIRD HOSPITAL
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