Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

A method of pig icsi operation

A sperm and lipase technology, applied in the field of pig ICSI operation, can solve problems such as hindering sperm, affecting sperm nucleus depolymerization, affecting oocyte activation, etc., and achieves the effect of increasing the formation rate and reducing adhesion problems.

Active Publication Date: 2017-10-20
INST OF ANIMAL SCI OF CHINESE ACAD OF AGRI SCI
View PDF2 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, when PVP is injected into the cytoplasm together with the sperm, it will surround the sperm, which not only makes the sperm plasma membrane tough, affects the rupture and delays the calcium ion oscillation in the cytoplasm, affects the depolymerization of the sperm nucleus, but also prevents the sperm from releasing the oocyte to the cytoplasm Cell activating factor, which affects the activation of oocytes, which seriously affects the success rate of ICSI

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • A method of pig icsi operation
  • A method of pig icsi operation
  • A method of pig icsi operation

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0042] The preparation method of in vitro maturation medium (IVM): in vitro maturation medium is made up of solvent and solute, and solvent is pure water; Solute and its concentration in culture medium: Medium 199 powder (Gibco, 31100-035) 9.5g / L , 10% (volume fraction) PFF, 10ng / ml EGF, 0.5mg / ml FSH, 0.5mg / ml LH, 70ng / ml Cys.

[0043] The preparation method of the activation solution: the activation solution is composed of a solvent and a solute, and the solvent is pure water; the solute and its concentration in the culture medium: 0.3M D-Mannitol, 0.5mM Hepes, 1.0mM CaCl 2 2H 2 O, 0.1mM MgCl 2 ·6H 2 O.

[0044]The preparation method of PZM-3 culture fluid: PZM-3 culture fluid is made up of solvent and solute, and solvent is pure water (Singma); Solute and its concentration in culture fluid: 6.3115g / L NaCl, 0.7455g / L KCl, 0.0476g / L KH 2 PO 4 , 0.0986g / L MgSO 4 ·7H 2 O, 2.1061g / L NaCO 3 , 0.022g / L sodium pyruvate, 0.6166g / L CaC 2 o 4 ·5H 2 O, 0.5% L-Glutamine, 0.05...

Embodiment 1

[0048] Embodiment 1, the method for improving pig ICSI operating efficiency and effect detection

[0049] 1. Methods for Improving Operational Efficiency of ICSI in Pigs

[0050] Experimental materials: Pig ovaries were collected from local slaughterhouses, placed in normal saline (38°C) with double antibodies (penicillin and streptomycin), and transported back to the laboratory within three hours.

[0051] 1. Wash the retrieved ovaries three times with normal saline with double antibodies, use a 10ml syringe and a 18-gauge needle to extract the follicle fluid with a diameter of 3-6mm follicles, and pick out 3-5 layers of eggs with a suction pipette. Cumulus-oocyte complexes (COCs) of cumulus cells.

[0052] 2. Wash the COCs selected in step 1 twice with the egg washing solution (PVA-TL-HEPES), and after cleaning the impurities, select the cumulus cells to wrap 3-5 layers, and the cumulus cells to wrap the complete oocyte densely and evenly Cells were transferred into in vit...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention discloses a pig ICSI operation method. The method of the present invention comprises the following steps: (1) using lipase or a lipase solution to hydrolyze the sperm of the isolated animal semen, collecting the hydrolyzate, and obtaining the sperm for preparing ICSI fertilized eggs. (2) injecting the sperm for preparing ICSI fertilized eggs into isolated oocytes to obtain ICSI eggs; (3) electrically activating and culturing the ICSI eggs to obtain ICSI fertilized eggs. The method of the invention not only avoids the adverse effects caused by the addition of PVP on the depolymerization of pig sperm, the formation of male pronuclei, and the later development, but also causes damage to the plasma membrane, acrosome and other regions of pig sperm, which contributes to After ICSI operation, porcine sperm released oocyte activating factor, which increased the formation rate of male pronucleus and the blastocyst formation rate of embryos after fertilization.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for pig ICSI operation. Background technique [0002] ICSI (Intracytoplasmic sperm injection) is intracytoplasmic sperm injection technology, which is the second generation of "test-tube baby". This technology uses a microscopic operating system to inject a single sperm into an egg to fertilize it. Compared with mice and humans, the membrane surface of pig sperm contains more lipids, glycolipids and glycoproteins and the acrosome region is rich in phospholipids and cholesterol. Therefore, during the ICSI operation, pig sperm tends to adhere to the injection needle, thus increasing the difficulty of micromanipulation. [0003] At present, a high molecular substance polyvinylpyrrolidone (PVP) is usually added to the operating solution to reduce the adhesion of the sperm to the injection needle and facilitate the operation. However, when PVP is injected into the c...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/076C12N15/89A01K67/027
Inventor 李奎刘志国魏迎辉
Owner INST OF ANIMAL SCI OF CHINESE ACAD OF AGRI SCI
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com