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A Method for Rapid Isolation and Line Establishment of Chicken Gonad Primordial Germ Cells

A technology of primordial germ cells and gonads, which is applied in the field of in vitro separation, cultivation and line establishment of animal reproductive stem cells, which can solve the problems of low efficiency, long time for line establishment, low success rate of line establishment of primordial germ cells, etc., and achieve stability Efficiency effect

Active Publication Date: 2021-08-17
GUANGXI UNIV
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  • Summary
  • Abstract
  • Description
  • 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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  • A Method for Rapid Isolation and Line Establishment of Chicken Gonad Primordial Germ Cells
  • A Method for Rapid Isolation and Line Establishment of Chicken Gonad Primordial Germ Cells
  • A Method for Rapid Isolation and Line Establishment of Chicken Gonad Primordial Germ Cells

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Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0031] First, material

[0032] The eggs used in the experiment are derived from the white chicken, the three yellow chickens in Guangxi, and the east is used.

[0033] Second, the method

[0034] Preparation for Chicken Embry

[0035] The cultivated eggs were incubated in the incubator, incubated at 37.5 ° C, 70% humidity conditions, and used in seventh day.

[0036] Separation of original reproductive cells

[0037] Extrafine eggs of the incubation of 7 days to the sterile operation chamber, rubbed the egg surface with alcohol, with a sterile bend, and remove the 7D embryo age chicken embryo, placed in a cell petri dish in 100 mm, Meter microscope observation. The chicken embryo is dissected under the body optic microscope, and gently scrape the gonads from one end of the ultra-fine thread, transferred to a preheated 30 μL of PBS droplets, and then washed twice, and then 1 pair of gonads The group was transferred into a 500 μl of 0.05% pneazic (containing 0.53 mM EDTA), and th...

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Abstract

The invention discloses a method for rapidly separating and establishing a line of primordial germ cells of chicken gonads. The primordial germ cells are separated and purified from the gonads of 7-day-old chicken embryos, and the isolated primordial germ cells are combined with the feeder layer by culture insert. Methods: Using the optimized culture system, more than 1 million cells were expanded within 15 days to achieve rapid expansion and line establishment. The invention has the characteristics of high success rate, short time, stability and high efficiency, and can be amplified within 15 days to obtain 10 6 The above cells have laid a foundation for the preservation of poultry germplasm resources and the preparation of transgenic chickens.

Description

Technical field [0001] The present invention belongs to the field of in vitro isolation, cultivation, and construction technology of animal reproductive stem cells, and more particularly to a method of rapid separation and construction of chicken gland original reproductive cells. Background technique [0002] Reproductive stem cells are a pluripotent stem cell having self-renewal and differentiation capabilities. The reproductive stem cells began to appear in the early embryos, migrated to reproductive ridges, and finally constituted an important part of the animal reproductive organ. During the animal breeding breeding, reproductive stem cells play an important role, not only play an important role in early emotional development of animals, and the process of self-renewal and differentiation capabilities are also maintained in the process of generating a distribution after body development. In the event, the egg played a decisive role. Related research of reproductive stem cell...

Claims

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Application Information

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Patent Type & Authority Patents(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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