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Isolated culture method of primary hepatocytes of chick embryos

A technology of primary hepatocytes, separation and culture, applied in the biological field, can solve the problems of separation and pollution of chicken hepatocytes, and achieve the effects of excellent activity, saving time and cost, and high activity.

Pending Publication Date: 2021-10-29
JIANGXI AGRICULTURAL UNIVERSITY
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Problems solved by technology

[0004] The purpose of the embodiments of the present invention is to provide a method for isolating and culturing primary hepatocytes of chicken embryos, aiming to solve the problem of pollution in the separation process of chicken hepatocytes

Method used

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  • Isolated culture method of primary hepatocytes of chick embryos
  • Isolated culture method of primary hepatocytes of chick embryos
  • Isolated culture method of primary hepatocytes of chick embryos

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Embodiment 1

[0034] Step 1: Chicken breeding eggs are routinely disinfected and treated, and then placed in a rotary incubator at 37.5°C and 70% humidity to hatch to the age of 12 embryos, and the chicken embryos hatched to the age of 12 are soaked and disinfected in 75% ethanol and placed in a non-toxic environment. Bacteria operating table;

[0035] Step 2: Take out the chicken embryo in step 1 under aseptic conditions, decapitate the chicken embryo and place it in a 0.01 mol / L phosphate buffered saline solution pre-cooled at 4°C;

[0036] Step 3: After cleaning the chicken embryos treated in step 2 twice, the blood on the surface of the chicken embryos is washed away;

[0037] Step 4: Put the chicken embryos cleaned in Step 3 into a sterile petri dish, use sterile ophthalmic forceps and ophthalmic scissors to remove the liver by laparotomy, and soak it in 4°C pre-cooled phosphate buffered saline solution;

[0038] Step 5: Cut the liver in step 4 into 1 mm 3 Small pieces of the same si...

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Abstract

The invention is applicable to the technical field of biology and provides an isolated culture method of primary hepatocytes of chick embryos. The isolated culture method comprises the following steps of: step 1, immersing and disinfecting the chick embryos which are hatched to 12 days old through 75 percent ethanol and putting the chick embryos into a sterile operation platform; step 2, taking out the chick embryos in the step 1 under a sterile condition; after carrying out end breaking treatment on the chick embryos, putting the chick embryos into a 0.01mol / L phosphate buffer salt solution which is pre-cooled at 4 DEG C; and step 3, after cleaning the chick embryos treated in the step 2 for two times, washing blood on the surfaces of the chick embryos. The SPF-grade chick embryos are adopted and the separation of livers can be finished in the operation platform; the operation is simple and convenient and bacterial pollution is not easy to occur. According to the method, collagenase is used for carrying out tissue digestion on the livers; compared with a traditional trypsin digestion method, damages to cells are relatively small and the damages caused by the collagenase are reversible; the hepatocytes with relatively good bioactivity can be obtained.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to a method for isolating and culturing chicken embryo primary liver cells. Background technique [0002] At present, there are few methods for culturing primary hepatocytes from chicken embryos in China. Chicken embryo primary hepatocytes cultured in vitro can not only maintain many unique functions of the liver, but also have good reproducibility of in vitro experiments, so they have been increasingly used to study the biological and metabolic functions of poultry liver middle. [0003] However, both mammalian and poultry primary isolated hepatocytes have problems such as high requirements for in vitro culture, short survival time, low purity, and difficult proliferation, especially the pollution that occurs during the isolation process of chicken hepatocytes. Contents of the invention [0004] The purpose of the embodiments of the present invention is to provide a method for isol...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071C12N5/073
CPCC12N5/067C12N5/0603C12N2509/00C12N2509/10C12N2500/25C12N2501/39
Inventor 杨帆幸程鸿曹华斌胡国良张彩英彭俊俊
Owner JIANGXI AGRICULTURAL UNIVERSITY
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