Application of cryoprotectant in organ and tissue cryopreservation

A cryopreservation solution and cryopreservation technology, applied in the field of biomedical materials, can solve the problems of affecting the survival rate of cryopreserved objects, safety function expression, cytotoxic side effects, and high toxicity of reagents.

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, such as: DMSO, lead to toxic side effects of cryopreservation reagents on cells, seriously affecting the survival rate and even (offspring) safety of cryopreservation objects after recovery and the functional expression
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 a problem of high reagent toxicity

Method used

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  • Application of cryoprotectant in organ and tissue cryopreservation
  • Application of cryoprotectant in organ and tissue cryopreservation
  • Application of cryoprotectant in organ and tissue cryopreservation

Examples

Experimental program
Comparison scheme
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preparation example Construction

[0112] 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.

[0113] The experimental methods used in the following examples are conventional methods unless otherwise specified; the reagents and materials used in the following examples can be obtained from commercial sources unless otherwise specified.

[0114] 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%.

[0115] In the cryopreservation solution in the embodiment of the present invention...

Embodiment 1

[0117] Example 1: Cryopreservation of Oocytes and Embryos

[0118] 1. Preparation of cryopreservation solution: prepare cryopreservation solution according to the following formula

[0119] Cryopreservation solution A:

[0120] Each 100mL contains the following components:

[0121] 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

[0122] 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...

Embodiment 2

[0163] Example 2: Stem Cell Cryopreservation

[0164] Cryopreservation solution C1:

[0165] Each 100ml contains the following components:

[0166] substance Dosage PVA (g) 1.0 Ethylene glycol (mL) 10 Sucrose (mol L -1 )

[0167] Cryopreservation solution D:

[0168] Each 100ml contains the following components:

[0169] substance Dosage PVA (g) 2.0 Ethylene glycol (mL) 10 Sucrose (mol L -1 )

[0170] Cryopreservation solution F:

[0171] Each 100ml contains the following components:

[0172] substance Dosage Poly-L-arginine (g, degree of polymerization is 8) 4.0 PVA (g) 1.0 Ethylene glycol (ml) 10 Sucrose (mol L -1 )

[0173] The preparation method of the cryopreservation solution is the same as in Example 1.

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Abstract

The invention discloses an application of a cryoprotectant in organ and tissue cryopreservation. The cryoprotectant contains PVA, ethylene glycol and sucrose. PVA is taken as a main cryoprotectant component of the cryoprotectant, the DMSO content and toxicity are low, and the cryoprotectant has high preservation efficiency when being applied to ovarian tissues or organs. The cryoprotectant is simple in composition and can be widely applied to cryopreservation of various cells, tissues and organs; the raw materials are easily available, and the cost is low.

Description

[0001] This application claims the priority of the previous application filed with the State Intellectual Property Office of China on April 9, 2019 with the patent application number 201910282416.X and the title of the invention "a PVA-based cryopreservation solution and its preparation method". The entirety of this prior application is incorporated by reference into the present application. technical field [0002] The invention belongs to the technical field of biomedical materials, and in particular relates to the application of a cryopreservation solution in organ and tissue cryopreservation. Background technique [0003] 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...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N1/02
CPCA01N1/0221
Inventor 严杰乔杰闫丽盈李蓉王健君金晟琳
Owner PEKING UNIV THIRD HOSPITAL
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