Duck embryo primary hepatocyte separation culture method

A technology for the isolation and cultivation of primary hepatocytes, which is applied in the field of isolation and cultivation of primary hepatocytes from duck embryos, can solve the problems of complex operation, high cost, low survival rate and purity of extracted cells, and achieve reduced contact concentration, full digestion, Effects of increasing separation cost and operational complexity

Active Publication Date: 2022-01-21
FEED RESEARCH INSTITUTE CHINESE ACADEMY OF AGRICULTURAL SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] In order to solve the problems of complex operation, high cost, low survival rate and purity of extracted cells in the prior art, the present invention is proposed and completed

Method used

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  • Duck embryo primary hepatocyte separation culture method
  • Duck embryo primary hepatocyte separation culture method
  • Duck embryo primary hepatocyte separation culture method

Examples

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Embodiment 1

[0043] Example 1 Isolation and Culture of Duck Embryo Primary Hepatocytes

[0044] 1. Isolation and cultivation

[0045] 1. Wipe and disinfect the shell of the duck embryo with iodophor, and spray and disinfect the shell of the duck embryo with 75% alcohol;

[0046] 2. Under aseptic conditions, break the shell from the end of the air chamber, open the eggshell membrane of the duck embryo with sterile tweezers, and take out the duck embryo;

[0047] 3. With the cooperation of surgical scissors and tweezers, the duck embryo is dissected to separate the duck embryo liver and remove the gallbladder, and the obtained liver is soaked and washed in D-Hank’s buffer;

[0048] 4. The scalpel and tweezers cooperate to remove the loose connective tissue and liver capsule around the liver, and soak the obtained small pieces of liver tissue in D-Hank’s buffer droplets. The soaking and washing process is repeated three times;

[0049] 5. Multi-step digestion, first soak the liver tissue in...

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Abstract

The invention relates to the field of cell culture, in particular to a duck embryo primary hepatocyte separation culture method. The method comprises the following steps: disinfecting shells of duck embryos, and taking 13-15-day-old duck embryos under an aseptic condition; separating the liver of the duck embryo, removing gall bladder, and soaking and washing the obtained liver in a D-Hank's buffer solution; removing loose connective tissues and liver envelopes adhered to the periphery of the liver, and soaking and washing the obtained small pieces of liver tissues in D-Hank's buffer solution; performing multi-step digestion; and re-suspending the obtained cells, sieving the cells with a 70-[mu]m cell sieve, and inoculating the cells into a culture bottle for culture. According to the invention, a simple, economical and rapid duck embryo primary hepatocyte separation method is established from the selection of the day age of the duck embryo, the combination of a mechanical method and an enzyme digestion method for tissues and the like.

Description

technical field [0001] The invention relates to the field of cell culture, in particular to a method for isolating and culturing duck embryo primary liver cells. Background technique [0002] The liver is an important internal organ for the metabolism of the animal body. It is the main place for the metabolism of carbohydrates, fats and proteins, and plays an important role in regulating the body's metabolism and maintaining the body's metabolic balance. Meat ducks and other poultry animals are different from mammals. Their adipocytes have limited ability to synthesize fat. The fat mainly comes from liver synthesis and intestinal absorption. Therefore, meat duck liver plays an extremely important role in the regulation of lipid metabolism. Isolating and culturing duck Hepatocytes provide a good basis for exploring the lipid metabolism and regulation mechanism of meat ducks. [0003] Liver cells are divided into hepatic parenchymal cells and hepatic non-parenchymal cells. Am...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071
CPCC12N5/067C12N2509/10C12N2509/00
Inventor 闻治国赵璐璐杨培龙蔡红英刘婧吴永保王帮磊
Owner FEED RESEARCH INSTITUTE CHINESE ACADEMY OF AGRICULTURAL SCIENCES
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