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Method for slaking oocyte transparent tape

An oocyte and zona pellucida technology, applied in the biological field, can solve the problems of incomplete dissolution of the zona pellucida, high cost, easy lysis and death of blastomeres, etc., and achieve the effects of not easily damaging cells, low cost, and short digestion time.

Inactive Publication Date: 2007-12-05
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The acidification method (Tyrode's, pH 2.5) has a fast action time, but requires rapid operation. The blastomeres obtained by the time is too long are easy to lyse and die, and the zona pellucida cannot be completely dissolved when the time is not enough; There is no adverse effect, but the processing conditions and time must be mastered, and the cost of using enzymes to digest the zona pellucida is higher, $182.7 per gram of trypsin
Therefore, the above methods have disadvantages

Method used

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  • Method for slaking oocyte transparent tape
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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0015] Example 1 Digestion of oocyte zona pellucida with glutathione

[0016] Preparation of glutathione: Weigh 1.00 g of reduced glutathione and dissolve it in 5 ml of sterilized physiological saline, divide into small tubes and store in a -20°C refrigerator for later use.

[0017] Operation: Thaw the frozen glutathione solution, then place the oocytes in the glutathione solution, digest for one minute and take out.

Embodiment 2

[0018] Example 2 Digestion of oocyte zona pellucida with glutathione

[0019] The oocytes were placed in freshly prepared 0.1g / mL glutathione solution, digested for 7 minutes and removed.

Embodiment 3

[0020] Example 3 Digestion of oocyte zona pellucida with glutathione

[0021] The oocytes were placed in freshly prepared 0.3g / mL glutathione solution, digested for 0.5 min and then removed.

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PUM

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Abstract

The present invention relates to method of digesting the zona pellucida of oocyte. The method is to set oocyte inside glutathione solution for digesting. The method of digesting the zona pellucida of oocyte in glutathione solution has simple operation, short digesting period, less damage to cell and low cost.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to a method for digesting oocyte zona pellucida. Background technique [0002] The zona pellucida is a homogeneous and distinct semipermeable membrane, the intercellular substance formed by follicle cells and oocytes, which can be dissolved by proteolytic enzymes such as trypsin and chymotrypsin. Digestion of zona pellucida is required to obtain blastomeres when mammalian blastomeres are isolated or when embryonic cells are used as donor nuclei for cloning. [0003] At present, there are three main methods for obtaining blastomeres: one is to digest the zona pellucida with protease, and then use microtubes to disperse the embryos into individual blastomeres; blastomeres; the third is to digest the zona pellucida with acidification. The acidification method (Tyrode's, pH 2.5) has a fast action time, but requires rapid operation. The blastomeres obtained by the time is too long are easy...

Claims

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Application Information

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IPC IPC(8): C12N5/06C12N5/08C12N5/075
Inventor 田见晖成文敏安磊
Owner CHINA AGRI UNIV
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