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

A method for improving the efficiency of porcine somatic cell nuclear transfer

A technology of culture medium and embryo culture medium, applied in botany equipment and methods, animal cells, vertebrate cells, etc., can solve the problem of low overall efficiency of pig somatic cell cloning, increase the number of blastocyst cells, and improve the overall efficiency , the effect of increasing blastocyst rate

Active Publication Date: 2019-11-29
南宁壮博生物科技有限公司
View PDF4 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although after two decades of development, cloning technology has been relatively mature, and there is a set of overall technologies including oocyte collection, culture, donor cell isolation, culture, egg activation, embryo in vitro culture and embryo transfer, but so far, The overall efficiency of pig somatic cell cloning is still very low, about 1%

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 for improving the efficiency of porcine somatic cell nuclear transfer
  • A method for improving the efficiency of porcine somatic cell nuclear transfer
  • A method for improving the efficiency of porcine somatic cell nuclear transfer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Embodiment 1, pig somatic cell nuclear transfer

[0044] 1. In vitro maturation culture of porcine oocytes

[0045] Take the pig ovary from the slaughterhouse, wash it three times with normal saline containing 100U / mL penicillin and 100U / mL streptomycin, and then use a 10mL disposable syringe and a 18-gauge needle to extract follicles with a diameter of 3-6mm Cumulus-oocyte complexes (COCs) wrapped with 3-5 layers of cumulus cells were selected from the liquid. COCs were rinsed twice with normal saline containing 100U / mL penicillin and 100U / mL streptomycin, then transferred to IVM solution, at 39°C, saturated humidity, 5% CO 2 Cultured in the incubator for 42h.

[0046] 2. Isolation and culture of porcine fetal fibroblasts

[0047] Pig fetal fibroblasts were prepared according to the following steps in turn:

[0048] 1. Take out the fetus from the uterus of pregnant sows at gestation day 35, wash with DPBS buffer solution containing 100U / mL penicillin and 100U / mL st...

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 method for improving the pig cell nucleus transplantation efficiency. An embryo culture solution is provided and contains 50 nM of BIX-01294 and 0.5 nM of chaetocin. The embryo culture solution can be used for culturing a reconstructed embryo and / or preparing a kit used for culturing the reconstructed embryo. BIX-01294 and chaetocin with certain concentration are added to a PZM-3 culture solution for combined treatment of an early cloned embryo, the blastocyst rate and the blastocyst cell number of the cloned embryo can be remarkably increased, the development efficiency of the cloned embryo is remarkably improved, and the method has a better effect than single use of BIX-01294 or chaetocin for treatment. The method has great significance for improving the overall efficiency of a pig cell nucleus transplantation technology.

Description

technical field [0001] The invention relates to a method for improving the efficiency of pig somatic cell nuclear transplantation. Background technique [0002] The technology of somatic cell nuclear transfer was born in 1997, and it was first successfully used in sheep. The main method is to transfer fully differentiated donor cells into enucleated oocytes to form reconstructed embryos through micromanipulation and cell fusion technology, and then activate the reconstructed embryos for culture and transplantation, and finally obtain a complete individual. Because the genotype of the obtained individual is exactly the same as that of the donor cell, this technique is also called somatic cell cloning technique. Due to its unique technical advantages, somatic cell cloning technology has great application value in animal breeding, disease model construction, endangered animal protection, therapeutic cloning, and pharmaceutical protein production. Since the birth of the first ...

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): C12N15/877C12N5/071
CPCC12N5/0602C12N15/8778C12N2500/30
Inventor 刘志国李奎牟玉莲郑新民毕延震
Owner 南宁壮博生物科技有限公司
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