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

Construction and application method of cynoglossus semilaevis pseudo male gonad cell line

A semi-smooth tongue sole and construction method technology, applied in artificial cell constructs, microbial-based methods, animal cells, etc., can solve the problems that cells cannot migrate out of tissue blocks, have not been reported, and the number of cells is small

Inactive Publication Date: 2014-11-19
YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
View PDF2 Cites 4 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The traditional cell culture method is not suitable for the establishment of the gonad tissue cell line of pseudomale tongue sole. For example, the number of cells obtained after treating the tissue with enzyme digestion method is very small, and the primary culture cannot be started; while the traditional tissue block method starts the primary culture, the tissue The block is difficult to adhere to and there is too much debris in the bottom of the culture flask. Cells cannot migrate out of the tissue block
The establishment of gonad tissue cell lines of flounder and flounder has always been a difficult point in fish cell culture, and so far there has been no report on the gonad tissue cell lines of pseudo-male fishes

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
  • Construction and application method of cynoglossus semilaevis pseudo male gonad cell line
  • Construction and application method of cynoglossus semilaevis pseudo male gonad cell line
  • Construction and application method of cynoglossus semilaevis pseudo male gonad cell line

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Healthy semi-smooth tongue sole was sterilized in 70% ethanol for 1 minute, the pseudo-male gonad tissue was removed in a sterile ultra-clean bench, the outermost membrane of the pseudo-male gonad tissue was peeled off, and the tissue was cut in 5% FBS-DMEM / F12 complete culture medium. into 1mm 3 Rinse 5 times with PBS, suspend the tissue pieces with 1ml serum-free DMEM / F12 medium, inoculate evenly to 25cm 2 Cell culture flasks were placed upside down in a biochemical incubator.

[0041] Prepare 100ml of special culture medium, take 80ml of DMEM / F12 liquid medium, add 20ml of fetal bovine serum, 10000IU of penicillin, 10mg of streptomycin, 1mg of tetracycline hydrochloride, 0.5μg of human recombinant basic fibroblast growth factor (bFGF), 2μg Insulin-like cell growth factor Ⅰ (IGF-1), 1000IU human chorionic gonadotropin (HCG) and 8000IU pregnant horse serum gonadotropin (PMSG).

[0042] After 6 hours, add 2ml of special culture medium, and turn the cell culture bottle...

Embodiment 2

[0045] Healthy semi-smooth tongue sole was sterilized in 70% ethanol for 1 minute, the pseudo-male gonad tissue was removed in a sterile ultra-clean bench, the outermost membrane of the pseudo-male gonad tissue was peeled off, and the tissue was cut in 5% FBS-DMEM / F12 complete culture medium. into 1mm 3 Rinse 6 times with PBS, suspend the tissue pieces with 1ml serum-free DMEM / F12 medium, inoculate evenly to 25cm 2 Cell culture flasks were placed upside down in a biochemical incubator.

[0046] Prepare 100ml of special culture medium, take 80ml of DMEM / F12 liquid medium, add 20ml of fetal bovine serum, 10000IU of penicillin, 10mg of streptomycin, 3mg of tetracycline hydrochloride, 0.8μg of human recombinant basic fibroblast growth factor (bFGF), 3μg Insulin-like cell growth factor Ⅰ (IGF-1), 1500IU human chorionic gonadotropin (HCG) and 9000IU pregnant horse serum gonadotropin (PMSG).

[0047] After 7 hours, add 2ml of special culture medium, and turn the cell culture bottle...

Embodiment 3

[0051] Healthy semi-smooth tongue sole was sterilized in 70% ethanol for 1 minute, the pseudo-male gonad tissue was removed in a sterile ultra-clean bench, the outermost membrane of the pseudo-male gonad tissue was peeled off, and the tissue was cut in 5% FBS-DMEM / F12 complete culture medium. into 1mm 3 Rinse 7 times with PBS, suspend the tissue pieces with 1ml serum-free DMEM / F12 medium, inoculate evenly to 25cm 2 Cell culture flasks were placed upside down in a biochemical incubator.

[0052] Prepare 100ml of special culture medium, take 80ml of DMEM / F12l liquid medium, add 20ml of fetal bovine serum, 10000IU of penicillin, 10mg of streptomycin, 5mg of tetracycline hydrochloride, 1μg of human recombinant basic fibroblast growth factor (bFGF), 4μg of Insulin-like cell growth factor Ⅰ (IGF-1), 10000IU human chorionic gonadotropin (HCG) and 2000IU pregnant horse serum gonadotropin (PMSG).

[0053] After 8 hours, add 2ml of special culture medium, and turn the cell culture bot...

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 aims to provide a construction method of a cynoglossus semilaevis pseudo male gonad cell line, thereby establishing the cynoglossus semilaevis pseudo male gonad cell line and providing a basis of researching sex differentiation and sex reversal regulatory mechanism of cynoglossus semilaevis. In the construction method, the cynoglossus semilaevis pseudo male gonad cell line is successfully established by employing cynoglossus semilaevis pseudo male gonad tissue. Continuous passage of the established cell line can be achieved. By means of the method, the passage cells can be generated for more than 60 generations and a large amount of cynoglossus semilaevis pseudo male gonad cells can be provided. The method can be widely applied in researches of exogenous gene, transfection of plasmids and functions of sex relative genes, can be applied in researches of breeding of common viruses in fish and a cooperative action mechanism of cells, and can also be applied in detection of environmental pollutants. The method has an important practical application value.

Description

technical field [0001] The invention belongs to the technical field of seawater fish cell culture, and in particular relates to a construction and application method of a seawater fish Cynoglossus semilaevis pseudomale gonad tissue cell line. Background technique [0002] Half-smooth tongue sole (Cynoglossus semilaevis) is an economic seawater fish in the northern coast of my country. There is a significant difference in growth rate between male and female individuals, and the growth rate of female fish is 2-4 times that of male fish. Studies have shown that tongue sole belongs to the ZW / ZZ sex determination mechanism, and the female fish has a heteromorphic W chromosome. In addition, research has found that there is a phenomenon of natural sex reversal in half-smooth tongue sole. The genetic sex of natural sex-reversed fish is female, but the biological sex is male, and its chromosomes are heterozygous (ZW type), which is called pseudo-male fish. According to the detection...

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): C12N5/071C12R1/91
Inventor 孙爱陈松林沙珍霞王娜
Owner YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES 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