Female chicken primordial germ cell expansion in vitro culture system and application thereof

A technology of primordial germ cells and culture system, which is applied in the field of in vitro expansion and culture system of primordial germ cells of female chickens, and can solve problems such as inapplicability

Active Publication Date: 2021-03-26
ANIMAL SCI RES INST GUANGDONG ACADEMY OF AGRI SCI
View PDF4 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, this type of in vitro culture system is more suitable for male chicken primordial germ cells, not for female chicken primordial germ cells

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
  • Female chicken primordial germ cell expansion in vitro culture system and application thereof
  • Female chicken primordial germ cell expansion in vitro culture system and application thereof
  • Female chicken primordial germ cell expansion in vitro culture system and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] (1) Preparation of aromatase culture medium

[0032] Find the sequence of the aromatase gene CYP19A1 (ENSGALG00000013294) on the ENSEMBL website, design primers (upstream: 5'-AGAAAACCTACTCAGA-3', downstream: 5'-GGCTTACAATAATGGTAG-3'), and use the gonad cDNA as a template to amplify the gene fragment. Use the In-Fusion kit (In-Fusion HD Cloning Kit, TaKaRa Company) to construct a vector with the target DNA, transform Escherichia coli DH5α (TaKaRa Company), pick positive clones, and send them to be sequenced to determine the sequence and expand the culture. Toxin plasmid extraction kit (Endo-free Plasmid mimi Kit II, OMEGA) was used to extract positive clone plasmid p-EGFP-CYP. Recovery of DF1 cells ( CRL-12203 TM , ATCC company) to 10cm cell culture dish (Thermo company), 37 ℃, 5% CO 2 Culture under conditions (the culture system is KO-DMEM (Gibco Company) with a volume ratio of 90%, fetal bovine serum (Gibco Company) with a volume ratio of 10%, and when the cells g...

Embodiment 2

[0041] The eggs of Huiyang bearded chickens (from the original breeding farm of the Institute of Animal Science, Guangdong Academy of Agricultural Sciences) were incubated for 5 to 7 days, and the chicken primordial germ cells were isolated from the gonad tissue of the chicken, and DNA was extracted from the limbs for sex identification. After sex identification, the female chicken primordial germ cells were inoculated onto the culture system and the rat liver cell feeder layer treated with the radioactive source in step (3) of Example 1, at 37° C., 5% CO 2 Conditioned incubator culture. The culture system is: 52% (v / v) KO-DMEM, 30% (v / v) rat liver cell culture medium, 7.5% (v / v) fetal bovine serum, 2.5% (v / v) chicken serum, 1% (v / v) non-essential amino acids, 1% (v / v) GlutaMAXTM, 1% (v / v) pyruvate, 1% (v / v) β-mercaptoethanol, 1% (v / v) penicillin Streptomycin, 3% (v / v) aromatase broth, 4ng / mL rmSCF, 2.5ng / mL rhFGF. A total of 16 female chicken embryos were isolated this time...

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 female chicken primordial germ cell expansion in vitro culture system and application thereof. The culture system consists of 52-54% (v / v) of KO-DMEM, 30% (v / v) of rat hepatocyte culture liquid, 7.5% (v / v) of fetal calf serum, 2.5% (v / v) of chicken serum, 1% (v / v) of non-essential amino acid, 1% (v / v) of GlutaMAXTM, 1% (v / v) of pyruvic acid, 1% (v / v) of [beta]-mercaptoethanol, 1% (v / v) of penicillin-streptomycin, 1-3% (v / v) of aromatizing enzyme culture liquid, 4ng / mL of rmSCF and 2.5ng / mL of rhFGF. The highest success rate for the culture system to amplify primordialgerm cells can be 37.5%, and in addition, the undifferentiation state and the good biological characteristics of the female chicken primordial germ cell can be kept.

Description

technical field [0001] The invention relates to the technical field of bioengineering, in particular to a female chicken primordial germ cell culture system in vitro and its application. Background technique [0002] The protection of poultry resources is mainly based on the conservation of living species, but the cost of feeding, site, and labor for the preservation of living species is high, and most of the conservation populations are temporarily unavailable. Therefore, it is necessary to develop other conservation technologies. Primordial germ cells (PGCs) originate from the epiblast and are the early cells that eventually form male or female germ cells, which can pass genes from the previous generation to the next generation. The isolation, cultivation and preservation of primordial germ cells is expected to realize the function of in vitro conservation of poultry resources, and if the primordial germ cells can be genetically manipulated, the sperm or eggs generated by ...

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): C12N5/071C12N15/53C12N15/70C12N9/02
CPCC12N5/0683C12N9/0071C12Y114/14C12N15/70C12N5/067C12N2500/84C12N2500/32C12N2500/44C12N2501/125C12N2501/119C12N2502/14C12N2509/00
Inventor 邹娴罗成龙王劼瞿浩舒鼎铭王艳吕晓慧
Owner ANIMAL SCI RES INST GUANGDONG ACADEMY 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