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Method for separating, culturing and identifying primary hepatic cells of livestock and fowl

A primary liver cell, livestock and poultry technology, applied in the biological field, can solve the problems of low vitality, low yield and purity of primary liver cells, cumbersome process, etc., and achieve less damage, easy adhesion, and good activity Effect

Inactive Publication Date: 2012-08-22
SHAOGUAN COLLEGE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the above separation methods have certain limitations. The main manifestations are: on the one hand, the yield and purity of primary hepatocytes are not high, their vitality is not strong, and the process is cumbersome; Research Provides Biological Materials

Method used

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  • Method for separating, culturing and identifying primary hepatic cells of livestock and fowl
  • Method for separating, culturing and identifying primary hepatic cells of livestock and fowl

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] 1. Isolation and purification of piglet liver cells

[0058] Select 1 7-day-old healthy Landrace piglet, drink water ad libitum and fast overnight before operation, intraperitoneally inject pentobarbital sodium (50mg / kg) for anesthesia;

[0059] Use gauze soaked in 75% alcohol to wipe the whole body of the piglet, place the anesthetized piglet in the supine position on the small animal operating table (the operating table is placed on a large porcelain plate), and transfer it to the ultra-clean workbench;

[0060] Open the abdominal cavity under aseptic conditions, gently pull the intestinal tract to the right side of the body, expose the mesenteric vein, moderately ligate both ends of the inferior vena cava, and insert a cannula in the direction of the hepatic portal vein (the part of the infusion tube that removes the needle bluntly). The cannula is then tied tightly to the vein to create access;

[0061] Perform perfusion separation: the first step is pe...

Embodiment 2

[0079] 1. Isolation and purification of chicken primary hepatocytes

[0080] Select 1 healthy young Lingnan yellow chicken, fast for 3 hours before operation, inject 1750IU / kg dose of heparin sodium into the wing vein for anticoagulation, and intraperitoneally inject pentobarbital sodium (50mg / kg) to anesthetize the animal;

[0081] Place the anesthetized chicken in the supine position on the small animal operating table (the operating table is placed on a large porcelain plate), wipe the whole body of the chicken with 75% alcohol, transfer it to the ultra-clean workbench, open the abdominal cavity under aseptic conditions, and gently The bowel is pulled to the left side of the body to expose the mesenteric veins, one of the retromesenteric veins that directly flows into the hepatic portal vein is cannulated (No. 5 scalp needle removes the needle), and the other vein that flows to the liver is ligated;

[0082] The first step is perfusion before digestion, which is di...

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Abstract

The invention relates to a method for separating, culturing and identifying primary hepatic cells of livestock and fowl, which integrates a chelation process and an in-situ two-step perfusion process. Ca<2+> and Mg<2+> in hepatic tissues of livestock and fowl are previously removed so that the hepatic tissues maximally release the primary hepatic cells; and the formula of additives of the culture fluid is adjusted to perfect the serum-free culture system and identify the glycogen of the obtained primary hepatic cells. The invention can effectively enhance the quantity, motility and purity of the separated primary hepatic cells of livestock and fowl, and is applicable to various animals; and the obtained primary hepatic cells have the advantages of fewer traumas, high wall attaching tendency and high activity, can be widely used for physiological function adjustment, metabolic diseases, and pharmacological and toxicological mechanisms of hepatic cells of livestock and fowl, can establish a virus infected cell model, and can be used for research in the fields of physics and chemistry, toxicant factor influence and the like.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a method for isolating, cultivating and identifying primary liver cells of various livestock and poultry animals. Background technique [0002] The liver is an important organ for the body to perform metabolic functions, deoxidize, store glycogen, synthesize secretory proteins and biotransform and other biological functions, and it is also the main place where drugs exert their toxic effects. Therefore, primary liver cells of livestock and poultry are used as biological materials in many in vitro studies, such as regulation of liver cell physiological functions, metabolic diseases, pharmacology, pathology and toxicology mechanisms, establishment of virus-infected cell models, and physical, chemical and The effects of toxic factors and other fields. [0003] It can be seen that the isolation, culture and identification of primary hepatocytes are the first steps to complete the above in...

Claims

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Application Information

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IPC IPC(8): C12N5/071C12Q1/00
Inventor 唐胜球江青艳朱晓彤董小英罗增福宾艳芳周桂炫方心灵
Owner SHAOGUAN COLLEGE
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