Livestock embryo vitrifying freeze process on metal surface

A vitrification freezing and metal surface technology, applied in the field of embryo biology, can solve the problems of weakening the antifreeze ability of embryos, embryo pollution, affecting the cooling speed, etc., and achieve the effect of promoting commercial application, improving economic benefits, and easy operation.

Inactive Publication Date: 2007-11-07
BEIJING JINXIU DADI AGRI CO LTD
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  • Abstract
  • Description
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  • Application Information

AI Technical Summary

Problems solved by technology

Due to the existence of microtubules in the microtube method, the freezing speed of embryos is reduced, which affects the efficiency of cryopreservation
The conventional operation of the micro-drop method is to immerse the embryo droplets directly into the liquid nitrogen. Since the liquid nitrogen may contain some pathogenic microorganisms, the direct contact of the embryo droplets with the liquid nitrogen will cause contamination of the embryos. In liquid nitrogen, due to the temperature difference, the liquid nitrogen around the embryo droplet is easy to vaporize and form an air mass, which affects the cooling rate
In the process of embryo freezing, the low temperature will cause certain damage to the cytoskeleton of the embryo, which weakens the antifreeze ability of the embryo to a certain extent.

Method used

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  • Livestock embryo vitrifying freeze process on metal surface

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Embodiment Construction

[0017] The present invention will be described in further detail below in conjunction with embodiment.

[0018] Take freezing ten bovine blastocysts as an example to illustrate the freezing process of the present invention:

[0019] 1. Preparation of freezing liquid:

[0020] 1.1 The manufacturers and article numbers of the reagents and drugs used are as follows

[0021] DPBS (Gibco, 15240-013)

[0022] DMSO (Sigma, D-5879)

[0023] Ethylene glycol (Sigma, E-9129)

[0024] Cytochalasin B (Sigma, C-6762)

[0025] Antibiotics (Gibco, 15240-062)

[0026] Sucrose (Sigma, S-1888)

[0027] Serum FBS (Hyclone, SH30070.03)

[0028] 1.2 DPBS base solution: Mix 39.5ml DPBS, 10ml FBS and 0.5ml antibiotics, filter and refrigerate at 4°C for later use

[0029] Equilibrium solution: Mix 10ml DPBS base solution and 50μg cytochalasin B, filter and refrigerate at 4°C for later use

[0030] VS1: Mix 8.5ml DPBS base solution, 0.75ml DMSO, 0.75ml ethylene glycol and 50μg cytochalasin B, ...

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Abstract

The present invention is livestock embryo vitrifying freeze process on metal surface, and belongs to the field of biotechnology. The livestock embryo vitrifying freeze process includes replacing freeze protectant for liquid inside and outside the embryo cell and fast freezing of the embryo liquid drop. During the fast freezing, the embryo liquid drop is dropped onto the exposed upper surface of one metal block with lower part soaked inside liquid nitrogen. The present invention can cool embryo in high speed while avoiding contamination of impurity in liquid nitrogen to the embryo. Besides, adding cytochalasin B as cytoskeleton protectant into the frozen liquid can raise the freezeproof capacity of the embryo and raise vitrification freezing effect. The present invention is used for freezing livestock embryo.

Description

technical field [0001] The invention relates to the technical field of embryo biology, in particular to a vitrification freezing method of livestock embryos. Background technique [0002] With the development of superovulation technology, in vitro fertilization, embryo transfer technology and animal cloning technology, livestock embryos have become commercial products. Embryo cryopreservation technology is extremely important for the commercial application of embryos and the promotion of cloning technology. Livestock embryo freezing technology has become a research hotspot in embryo engineering technology. In recent years, embryo cryopreservation technology, especially vitrification, has developed rapidly. Vitrification technology is the result of the combination of rapid freezing and high-concentration cryoprotectant. The high-concentration cryoprotectant forms a transparent glass-like solid during rapid cooling. During the cooling process, the viscosity is very high, but ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N1/02
Inventor 张家新
Owner BEIJING JINXIU DADI AGRI CO LTD
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