Immortalized dairy cow mammary epithelial cell line, construction method and applications thereof

A mammary epithelial cell, immortalization technology, applied in the field of immortalized cell lines, immortalized dairy cow mammary epithelial cell lines and their construction, can solve the problems of morphological differentiation aging, instability, insufficient number and types of mammary epithelial cell lines, etc. Achieve the effect of obvious cell boundary line, uniform cell shape, and solve the difficulty of culture

Active Publication Date: 2015-09-23
INST OF ANIMAL SCI OF CHINESE ACAD OF AGRI SCI
View PDF0 Cites 5 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

BMGH+HM does not have the polygonal morphology of typical epithelial cells, but has a long and thin shape with long protrusions, but can synthesize cytokeratin and desmosomal proteins; BMGE+H has typical epithelial cell morphology and can synthesize cytokeratin , but cannot form domes or bubble-like structures, and cannot synthesize vimentin
[0005] Although there are many reports on th

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
  • Immortalized dairy cow mammary epithelial cell line, construction method and applications thereof
  • Immortalized dairy cow mammary epithelial cell line, construction method and applications thereof
  • Immortalized dairy cow mammary epithelial cell line, construction method and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Example 1 Construction and Identification of Immortalized Cow Mammary Epithelial Cell Line

[0044] 1. Construction of Immortalized Cow Mammary Epithelial Cell Line

[0045] 1.1pBABE-Puro retroviral vector

[0046] The retrovirus plasmid and the pBABE-SV40-largeT antigen plasmid were both donated by teachers from the Institute of Zoology, Chinese Academy of Sciences. figure 1 This is the map of the pBABE-Puro Retroviral Vector retroviral vector used.

[0047] pBABE-puro-SV-40-large-T plasmid

[0048] Take out frozen DH5α competent cells (100 μL each) and thaw on ice, add 0.5 μL plasmid, mix well, place on ice for 30 minutes, incubate at 42°C for 60-90s, then place on ice for 2 minutes; add 37°C to warm 900 μL of SOC medium, shake the bacteria at 220 rpm at 37°C for 1 h; take 200 μL of the bacterial solution and spread it on the LB agar plate containing kanamycin, and cultivate it overnight at 37°C; pick 10 clones from the LB agar plate, add Cultivate overnight in LB...

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 an immortalized dairy cow mammary epithelial cell line, a construction method, and applications thereof. In the provided construction method, Chinese Holstein cow epithelial cells are taken as the host cells, retrovirus carriers with SV-40-large T antigen are used to infect host cells, and finally a strain of immortalized dairy cow mammary epithelial cell line that can perform stable passage is screened out. The microbial preservation number of the strain is CGMCC No.8707. The constructed immortalized dairy cow mammary epithelial cell line has a stable property, the cell morphology is similar to that of primary cells after 40 generations, and the cells grow faster than the primary cells, can characteristically express mammary epithelial cell specific proteins, and maintain the synthesis functions of milk fat and milk protein. The immortalized dairy cow mammary epithelial cell line can be used as a cell model which is widely used in the fields such as milk component synthesis, milk secretion mechanism, research on laticiferous cell apoptosis and mammary gland degradation mechanism, and the like, and can be used as a mammary gland bioreactor to produce bioactive proteins.

Description

technical field [0001] The invention relates to an immortalized cell line, in particular to an immortalized dairy cow mammary gland epithelial cell line and its construction method and application, and belongs to the field of construction and application of the immortalized dairy cow mammary gland epithelial cell line. Background technique [0002] Cow mammary gland epithelial cells have special functions of synthesizing and secreting milk, and can maintain their characteristics under in vitro culture conditions. The research on cow mammary epithelial cells has been more than half a century. Cow mammary gland epithelial cells can be used to study the expression of milk protein and milk fat genes and the mechanism of milk secretion. As a cell model of mammary gland bioreactor to produce valuable bioactive proteins, it is a research tool with great application value. However, there are great difficulties in the in vitro culture of dairy cow mammary epithelial cells, and the ...

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
IPC IPC(8): C12N5/10C12N15/867C12R1/91
Inventor 郑楠胡菡王加启高海娜代文婷
Owner INST OF ANIMAL SCI OF CHINESE ACAD OF AGRI SCI
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