Technique for freezing bare embryo

A cryopreservation and embryo technology, applied in the preservation of microorganisms, human or animal bodies, biochemical equipment and methods, etc., can solve the problems of low temperature resistance, damage to exposed cell groups, etc.

Inactive Publication Date: 2006-02-01
SHANDONG NEW HOPE LIUHE GROUP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] In order to solve the problem that the low temperature resistance of the embryo is reduced after the zona pellucida is removed, and at the same time, the penetration function of the freezing protective liquid is strengthened, causing damage to the exposed cell mass due to solute effects and physical damage during the freezing process, a new method is invented. A method of freezing naked embryos with ethylene glycol, sucrose, dextran, etc. as the main raw materials

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] 1. Dispensing: Take 2ml of ethylene glycol, 1ml of sucrose, 1ml of dextran, 1.5ml of serum albumin, 2.5ml of PBS, and 2ml of ultrapure water.

[0022] 2. Turn on the ultraviolet disinfection lamp of the sterile operating table, and operate after irradiating for 30 minutes. Make dextran into a 10% aqueous solution, then take 2ml of ethylene glycol, 1ml of sucrose, 1ml of dextran, 1.5ml of serum albumin, 2.5ml of PBS, and 2ml of ultrapure water, and put them into a small glass after high temperature disinfection bottle, mix well. After filtering with a bacterial filter, store at a constant temperature of 4°C for 5-7 days.

[0023] 3. The recovery rate of naked embryos frozen with this formula is 87% after thawing, and the success rate of transplantation is 42%.

Embodiment 2

[0025] 1. Dispensing: Take 3ml of ethylene glycol, 1ml of sucrose, 1ml of dextran, 1ml of serum albumin, 3ml of PBS, and 1ml of ultrapure water.

[0026] 2. Turn on the ultraviolet disinfection lamp of the sterile operating table, and operate after irradiating for 30 minutes. Make dextran into a 10% aqueous solution, then take 3ml of ethylene glycol, 1ml of sucrose, 1ml of dextran, 1ml of serum albumin, 3ml of PBS, and 1ml of ultrapure water, and put them into a small glass bottle after high temperature disinfection , mix well. After filtering with a bacterial filter, store at a constant temperature of 4°C for 5-7 days.

[0027] 3. The recovery rate of naked embryos frozen with this formula is 81% after thawing, and the transplantation success rate is 40%.

Embodiment 3

[0029] 1. Dispensing: Take 2ml of ethylene glycol, 2ml of sucrose, 1ml of dextran, 1ml of serum albumin, 3ml of PBS, and 1ml of ultrapure water.

[0030] 2. Turn on the ultraviolet disinfection lamp of the sterile operating table, and operate after irradiating for 30 minutes. Make dextran into a 10% aqueous solution, then take 2ml of ethylene glycol, 2ml of sucrose, 1ml of dextran, 1ml of serum albumin, 3ml of PBS, and 1ml of ultrapure water, and put them into small glass bottles after high temperature disinfection , mix well. After filtering with a bacterial filter, store at a constant temperature of 4°C for 5-7 days.

[0031] 3. The recovery rate of naked embryos frozen with this formula is 86% after thawing, and the success rate of transplantation is 40%.

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Abstract

A method for freezing the bare embryo features that its freezing liquid contains ethanediol (20-30%), cane sugar (10-20%), glucosan (10-15%), serum albumin (10-15%), PBS (10-30%) and ultra-pure water (10-20%). Its advantages are high activity of thawed embryo and high transplantation success rate.

Description

Technical field: [0001] The invention relates to the field of embryo transplantation and frozen embryo production, in particular to a freezing method for ultra-low temperature preservation after the embryo is removed from the zona pellucida. Background technique: [0002] At present, the gender identification technology of early embryos must obtain some embryonic cells for gene amplification, so as to determine the sex of the embryo and achieve the purpose of sex control. However, after the embryo is cut and sampled, the zona pellucida is destroyed. If the exposed cell mass is put back into another zona pellucida, the required equipment investment is very large, and it is inconvenient to carry, which is not conducive to industrial operation. Therefore, the use of unique freezing liquid and freezing technology can greatly reduce unnecessary investment and facilitate large-scale promotion. Invention content: [0003] In order to solve the problem that the low temperature re...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/08C12N5/06A01N1/02
Inventor 黄河李鑫
Owner SHANDONG NEW HOPE LIUHE GROUP
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