Method for separating and culturing primary chicken hepatocytes

A chicken hepatocyte isolation and culture technology, applied in the field of isolation and culture of primary chicken hepatocytes, can solve the problems of affecting the basic metabolism of liver cells, interfering with biotransformation, bioassay interference, etc., to reduce the risk of cell contamination and achieve good perfusion effect , the effect of long cultivation time

Inactive Publication Date: 2014-01-22
NANJING AGRICULTURAL UNIVERSITY
View PDF4 Cites 19 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the addition of serum to culture cells also has its disadvantages: First, the composition of serum is very complex and unclear. The use of serum in hepatocyte culture will affect the basic metabolism of hepatocytes and affect the research and production based on hepatocyte culture. Second, some biologically active substances in serum can poison liver cells and interfere

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Method for separating and culturing primary chicken hepatocytes
  • Method for separating and culturing primary chicken hepatocytes
  • Method for separating and culturing primary chicken hepatocytes

Examples

Experimental program
Comparison scheme
Effect test

Example Embodiment

[0026] Example 1

[0027] In this example, a 30-day-old 1.5kg healthy white feather broiler was used to isolate and culture hepatocytes. Before taking the liver, fasting for 3 hours, intravenous injection of 2250IU of heparin sodium under the wings, 5 minutes later, the brain was sacrificed, and the chickens were soaked with 0.1% neocervical solution for disinfection, ensuring that the skin and coat were wetted by the solution. Then bring the chicken into a sterile room, fix it on its back, and wipe the abdominal skin with alcohol cotton. Open the abdominal cavity, first ligate the common mesenteric vein, pancreaticoduodenal vein, and proventriculus splenic vein of the right hepatic portal vein into the right liver, and then ligate the gastroabdominal vein, left gastric vein, and posterior glandular gastric vein into the left hepatic portal vein , And finally ligate the posterior vena cava and common iliac vein. Cut the blood vessels, take out the liver, and transfer to the ult...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention discloses a method for separating and culturing primary chicken hepatocytes. According to the method, EGTA (Ethylene Glycol Tetraacetic Acid) with low hepatotoxicity is added into a perfusate A to loosen the junction between hepatocytes, so that the dispersion degree of the to-be-separated hepatocytes is increased; double resistance is added into the perfusate A, a perfusate B and D-Hanks flushing fluid, so that the pollution is further prevented; and circulating perfusion digestion is adopted during digestion perfusion, so that the perfusion cost is greatly reduced. A serum-free cultural method of the chicken hepatocytes is established; compared with a traditional serum cultural method, the method disclosed by the invention has the same effect on the aspects of cell quantity, activity, function and the like, so that the problems caused by the existence of serum are solved and a foundation is laid for the substance metabolism study and biological product preparation on the basis of the chicken hepatocyte culture.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for isolating and culturing primary chicken hepatocytes. Background technique [0002] Compared with other in vitro and in vivo experimental methods, the use of primary hepatocytes for in vitro experiments has its own significant advantages. The primary cultured hepatocytes are not affected by the complex neuroendocrine system in the body, and can maintain the specific functions of the hepatocytes and maintain the responsiveness to some hormones. Therefore, the primary cultured hepatocytes are ideal for studying liver substance metabolism and An excellent model of its regulatory mechanism. [0003] The isolation of hepatocytes was initially performed by the isolated perfusion method. By 1975, Seglen successfully isolated rat hepatocytes with high activity rate by in situ two-step collagenase perfusion method. In 1992, Fraslin et al. improved Seglen's in situ two-step perfu...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
IPC IPC(8): C12N5/071
Inventor 黄克和车超平杨玉澜潘翠玲陈兴祥
Owner NANJING AGRICULTURAL UNIVERSITY
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap