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Blastocyst vitrification refrigerating fluid and refrigerating method

A technology of vitrification and freezing liquid, which is applied in the field of animal embryo engineering, can solve the problems of long time-consuming effect and large difference in effect, and achieve the effect of short time-consuming and improving freezing efficiency

Inactive Publication Date: 2020-06-16
佛山辅康生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, different cryoprotectants and their combinations have different effects in different embryo freezing
Moreover, the general effect of the existing blastocyst freezing solution is relatively common and takes a long time (the pre-protection solution needs to act for 10-15 minutes, and the protection solution needs to act for 3-5 minutes)

Method used

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  • Blastocyst vitrification refrigerating fluid and refrigerating method
  • Blastocyst vitrification refrigerating fluid and refrigerating method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] This embodiment provides a blastocyst vitrification freezing solution, the preparation process is as follows:

[0041] 1. Prepare 10ml balance solution: add 0.5ml HSA to 9.5ml mHTF.

[0042] 2. Prepare 10ml freezing solution 1: Take 0.2g of PVP freeze-dried powder, 0.75ml of EG and 0.75ml of DMSO to 10ml with balance solution, dissolve completely and mix.

[0043] 3. Prepare 10ml freezing solution 2: Add 2.27g of trehalose, 0.2g of PVP freeze-dried powder, 1.5ml of EG, 1.5ml of DMSO and 0.12g of polysucrose, dilute to 10ml with a balance solution, completely dissolve and Mix well.

Embodiment 2

[0045] This embodiment provides a blastocyst vitrification freezing solution, the preparation process is as follows:

[0046] 1. Prepare 10ml balance solution: add 0.5ml HSA to 9.5ml mHTF.

[0047] 2. Prepare 10ml freezing solution 1: Take 0.1g of PVP freeze-dried powder, 1ml of EG and 1ml of DMSO to 10ml with balance solution, dissolve completely and mix.

[0048] 3. Prepare 10ml of freezing solution 2: Mix 2.27g of trehalose, 0.1g of PVP freeze-dried powder, 2ml of EG, 2ml of DMSO and 0.12g of polysucrose, dilute to 10ml with balance solution, dissolve and mix thoroughly .

Embodiment 3

[0050] This embodiment provides a blastocyst vitrification freezing solution, the preparation process is as follows:

[0051] 1. Prepare 10ml balance solution: add 0.5ml HSA to 9.5ml mHTF.

[0052] 2. Prepare 10ml freezing solution 1: Take 0.3g of PVP freeze-dried powder, 0.75ml of EG and 0.75ml of DMSO to 10ml with balance solution, dissolve completely and mix.

[0053] 3. Prepare 10ml freezing solution 2: Add 2.27g of trehalose, 0.3g of PVP freeze-dried powder, 1.5ml of EG, 1.5ml of DMSO and 0.12g of polysucrose, dilute to 10ml with a balance solution, completely dissolve and Mix well.

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Abstract

The invention relates to a blastocyst vitrification refrigerating fluid and a refrigerating method. The vitrification refrigerating fluid comprises an equilibrium liquid, a refrigerating fluid 1 and arefrigerating fluid 2; wherein the equilibrium liquid comprises a basic equilibrium liquid and a protein additive; the refrigerating fluid 1 comprises polyvinylpyrrolidone, ethylene glycol, dimethylsulfoxide and the equilibrium liquid; the refrigerating fluid 2 comprises polyvinylpyrrolidone, ethylene glycol, dimethyl sulfoxide, trehalose, polysucrose and the equilibrium liquid. The vitrification refrigerating fluid provided by the invention can significantly improve the freeze-thaw survival rate and the recruitment rate of blastocysts, and is short in time consumption.

Description

Technical field [0001] The invention relates to animal embryo engineering, in particular to a vitrification freezing liquid and freezing method for blastocysts. Background technique [0002] The vitrification technology is to cool the cell and its protective agent solution to the "glass transition temperature" at a sufficiently fast cooling rate to be solidified into a complete glass state, and store it at a low temperature in this glass state to avoid The formation of ice crystals reduces the freezing damage of ice crystals to cells and achieves the effect of cryoprotection. [0003] In recent years, the vitrification technology of blastocyst stage embryos has made significant progress, and the survival rate after freezing and thawing has been greatly improved. However, because blastocysts have undergone self-selection and elimination during the blastocyst culture period, the rate of available embryos is low, so there is a risk of no embryos to be transferred. How to further imp...

Claims

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Application Information

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IPC IPC(8): A01N1/02
CPCA01N1/0221
Inventor 姜永存胡威曾玉洁赵衡斌陈烨周星宇吴秀芝吴文林
Owner 佛山辅康生物科技有限公司
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