A kind of cryopreservation liquid and preparation method thereof

A cryopreservation liquid and solution technology, applied in the field of biomedical materials, can solve the problems of affecting the survival rate of cryopreservation objects, the safety of functional expression, the inability to effectively control the growth of ice crystals, and the lack of ice crystal growth, etc., to achieve high cell survival rate and Functional expression stability, excellent spreading performance, and convenient raw material sources

Active Publication Date: 2022-03-25
北京玳衲芈珂生物科技有限公司
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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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  • A kind of cryopreservation liquid and preparation method thereof
  • A kind of cryopreservation liquid and preparation method thereof
  • A kind of cryopreservation liquid and preparation method thereof

Examples

Experimental program
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preparation example Construction

[0074] The preparation method of the present invention will be described in further detail below with reference to specific examples. 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 implemented based on the above content of the present invention are covered within the intended protection scope of the present invention.

[0075] The experimental methods used in the following examples are conventional methods unless otherwise specified; the reagents, materials, etc. used in the following examples can be obtained from commercial sources unless otherwise specified.

[0076] The PVA used in the embodiment of the present invention has a syndiotacticity of 50%-55%, a molecular weight of 13-23 kDa, and a degree of hydrolysis of 98%.

[0077] In the embodiment of the present invention, the poly-L-proline polymerizat...

Embodiment 1

[0079] Example 1: Cryopreservation of oocytes and embryos

[0080] 1. Prepare cryopreservation solution: prepare cryopreservation solution according to the following formula

[0081] Cryopreservation Solution A:

[0082] Each 100mL contains the following components:

[0083] 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

[0084] 2.0 g of PVA was heated in a water bath at 80°C and dissolved in 25 mL 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; 17 g (0.05 mol) of sucrose (sucrose was frozen in The final concentration in the preservation solution is 0.5mol L -1 ) ultrasonically dissolved in 25 mL of DPBS, after the sucrose was completely dissolved, add 10 mL of ethylene glycol and 10 mL of DMSO to obtain solution 2. After solution...

Embodiment 2

[0128] Example 2: Stem cell cryopreservation

[0129] Cryopreservation solution C1:

[0130] Each 100ml contains the following components:

[0131] substance Dosage PVA(g) 1.0 Ethylene glycol (mL) 10 Sucrose (mol L -1 )

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

[0132] Cryopreservation Solution D:

[0133] Each 100ml contains the following components:

[0134] substance Dosage PVA(g) 2.0 Ethylene glycol (mL) 10 Sucrose (mol L -1 )

0.5 DPBS(mL) margin

[0135] Cryopreservation Solution F:

[0136] Each 100ml contains the following components:

[0137]

[0138]

[0139] The preparation method of cryopreservation solution is the same as that of Example 1.

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Abstract

The invention discloses a cryopreservation solution and a preparation method thereof, which contains PVA, ethylene glycol and sucrose. The cryopreservation solution uses PVA as the main cryoprotectant component, the DMSO content is low, and the toxicity is small. When it is used for the cryopreservation of cells or tissues, especially oocytes, it can reach the same level as the commercial cryopreservation solution (containing DMSO 15%). Equal or even higher cell survival rate and functional expression stability, with higher preservation efficiency. The cryopreservation solution of the invention has simple composition, convenient source of raw materials and low cost, and can be widely used in cryopreservation of various cells, tissues and organs.

Description

[0001] This application claims the priority of the prior application with the patent application number 201910282416.X submitted to the State Intellectual Property Office of China on April 9, 2019, and the invention name is "A PVA-based cryopreservation solution and its preparation method". The entire contents of this prior application are incorporated herein by reference. technical field [0002] The invention belongs to the technical field of biomedical materials, and in particular relates to a PVA-based cryopreservation solution and a preparation method thereof. Background technique [0003] Cryopreservation refers to the preservation of cells, tissues or organs under ultra-low temperature, which slows down or stops cell metabolism and division, and can continue to develop once the normal physiological temperature is restored. Since its inception, this technology has become one of the indispensable research methods in the field of natural sciences and has been widely used...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01N1/02
CPCA01N1/0221
Inventor 王健君金晟琳严杰乔杰闫丽盈李蓉
Owner 北京玳衲芈珂生物科技有限公司
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