A method for improving the in vitro development efficiency of pig cloned embryos

A high-efficiency technology for cloning embryos, applied in the field of reproduction or insemination, can solve the problems of low development efficiency of pig cloned embryos in vitro, and achieve the effects of increasing the number of cells at the blastocyst stage, increasing the division rate, and high blastocyst rate

Inactive Publication Date: 2015-12-02
SUN YAT SEN UNIV
View PDF3 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The technical problem to be solved by the present invention is to overcome the defect of low in vitro development efficiency of pig cloned embryos in the prior art, and provide a method for improving the in vitro development efficiency of pig cloned embryos

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 in vitro development efficiency of pig cloned embryos
  • A method for improving the in vitro development efficiency of pig cloned embryos
  • A method for improving the in vitro development efficiency of pig cloned embryos

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] 1. Materials

[0033] αMEM culture medium was purchased from LifeTechnologies; reagents were purchased from sigma; antibodies were purchased from eBioscience; hKlf4 (K, carrying human Klf4 gene sequence, Genbank: NM_004235.4), pLv-CMV-ZsGreen-hc-Myc (M, carrying human c-Myc gene sequence, Genbank: NM_002467.4), pLv-CMV-ZsGreen- hLin28 (L, carrying the human Lin28 gene sequence, Genbank: NM_024674.4), pLv-CMV-ZsGreen-hNanog (N, carrying the human Nanog gene sequence, Genbank: NM_024865.2), pLv-CMV-ZsGreen-hSox2 (S, Carrying the human Sox2 gene sequence, Genbank: NM_003106.3) was preserved by the Animal Genetic Engineering Laboratory of the School of Life Sciences, Sun Yat-sen University.

[0034] Two, test procedure: the whole flow diagram of the present invention sees as figure 1 shown.

[0035] S1. Isolation and purification of bone marrow mesenchymal stem cells

[0036] S11. Separation of bone marrow:

[0037] Extract 5~10mL of porcine tibial bone marrow, dilute...

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 in-vitro development efficiency of porcine cloned embryos. The method comprises the following steps of: performing molecular identification on bone marrow-derived mesenchymal stem cells, and establishing a method capable of identifying the purity of the cells by using a few cells through a 6-channel flow type technology; performing gene modification on the porcine bone marrow mesenchymal stem cells to obtain porcine bone marrow mesenchymal stem cells for expressing multiple reprogramming factors; and separating out positive porcine bone marrow mesenchymal stem cells by using a fluorescent flow type separation method, continuously culturing the positive porcine bone marrow mesenchymal stem cells for 3 to 7 days, performing nuclear transfer on the cultured cells serving as nuclear donor cells, and studying the influence of 4RFs-3days, 6RFs-3days, 4RFs-7days and 6RFs-7days pMSC serving as nuclear donor cells on the improvement of the development efficiency of the porcine cloned embryos. The results show that the 4RFs-7days pMSC can well promote fission of the cloned embryos and formation of blastospheres, the cloned embryos can form a homogeneous state with uniform cell quantity, and a foundation is laid for efficiently cloning adult high-quality pig varieties in large scale.

Description

technical field [0001] The invention relates to a method for improving the in vitro development efficiency of pig cloned embryos, in particular to a method for improving the in vitro development efficiency of pig cloned embryos by using reprogramming factors to genetically modify donor cells, and belongs to the field of reproduction or fertilization methods. Background technique [0002] Pigs have very similar organs to humans, as well as similar anatomical and physiological characteristics, and have gradually become biological model materials for regenerative medicine research. People are increasingly using nuclear transfer technology to produce pigs with certain genetic defects or modified genes as disease models to better simulate the clinical treatment of human diseases. In addition, in the actual production of pigs, some high-quality germplasm resources cannot give full play to the breeding advantages due to the limited number of years of use, and excellent individuals ...

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): C12N15/85C12N5/10C12N15/873
Inventor 宋振威丛佩清冀倩倩赵海静何祖勇陈瑶生
Owner SUN YAT SEN UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap