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Method for rapid separation and line establishment of chicken gonad primordial germ cells

A primordial germ cell and gonad technology, applied in germ cells, biochemical equipment and methods, embryonic cells, etc., can solve the problems of instability, inefficiency, and long line establishment time of the culture system, and achieve good gonad migration ability. , high success rate, short time effect

Active Publication Date: 2018-08-10
GUANGXI UNIV
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  • Abstract
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  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these methods have a low success rate (about 5%-30%) and a long time to establish a line (30-60d) to obtain primordial germ cells, and the culture system is unstable and inefficient.

Method used

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  • Method for rapid separation and line establishment of chicken gonad primordial germ cells
  • Method for rapid separation and line establishment of chicken gonad primordial germ cells
  • Method for rapid separation and line establishment of chicken gonad primordial germ cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0031] 1. Materials

[0032] The eggs used in the experiment were from white feather chicken, Guangxi Sanhuang chicken and Donglan black-bone chicken.

[0033] 2. Method

[0034] Preparation of chicken embryos

[0035] The fertilized eggs were hatched in the incubator at 37.5°C and 70% humidity, and were taken on the seventh day.

[0036] Isolation of primordial germ cells

[0037] Take out the fertilized eggs hatched for 7 days to the aseptic operation room, spray the surface of the eggs with alcohol, open the big end of the eggs with sterile curved tweezers, take out the 7d embryonic chicken embryos, place them in a 100mm cell culture dish, Observe under a stereomicroscope. Dissect the chicken embryo under a stereomicroscope, gently scrape off the gonads with one end of ultra-fine tweezers, transfer them to a pre-prepared 30 μL drop of PBS for washing, repeat the washing twice, and use one pair of gonads as a pair Transfer to a 500 μL centrifuge tube containing 20 μL 0...

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Abstract

The invention discloses a method for rapid separation and line establishment of chicken gonad primordial germ cells. The method comprises: separating and purifying primordial germ cells from the gonadof a 7-day-old chick embryo, and amplifying the separated primordial germ cells within 15 days by using a culture insert matched feeder layer method through an optimized culture system to obtain morethan one million cells so as to achieve rapid amplification and line establishment. According to the present invention, the method has characteristics of high success rate, short time, stability andhigh efficiency, can obtain more than 10<6> cells through the amplifying within in 15 d, and can establish the foundation for the poultry germplasm source conservation and preparation of transgenic chicken.

Description

technical field [0001] The invention belongs to the technical field of in vitro separation, cultivation and line establishment of animal reproductive stem cells, and in particular relates to a method for rapid separation and line establishment of chicken gonad primordial germ cells. Background technique [0002] Germ stem cells are a group of pluripotent stem cells capable of self-renewal and differentiation with the ability to produce gametes. Germ stem cells begin to appear in early embryos, migrate to the gonads to form gonads, and eventually constitute an important part of animal reproductive organs. In the process of animal reproduction and breeding, reproductive stem cells play an important role, not only in the early embryonic development of animals, but also in the process of producing gametes after the body matures, and the maintenance of its self-renewal ability and differentiation ability is also important for sperm Occurrence and ovum play a decisive role. Rele...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0735
CPCC12N5/0611C12N2500/30C12N2500/32C12N2500/38C12N2500/40C12N2500/44C12N2501/115C12N2501/91
Inventor 陆阳清谢龙陆振萍陈东阳
Owner GUANGXI UNIV
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