Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Preparation method of tissue-engineered hepatic model

A technology of tissue engineering and modeling, applied in biochemical equipment and methods, tissue culture, liver cells, etc., can solve the problems of maintaining liver structure and function, and achieve long-term drug exposure, high mitochondrial function, and increased expression levels.

Inactive Publication Date: 2020-01-14
GUANGDONG BOXI BIO TECH CO LTD
View PDF0 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The purpose of the present invention is to overcome the defect that the existing liver model cannot maintain the structure and function of the liver for a long time in vitro, so as to better perform functional tests such as drug metabolism and toxicity evaluation

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
  • Preparation method of tissue-engineered hepatic model
  • Preparation method of tissue-engineered hepatic model
  • Preparation method of tissue-engineered hepatic model

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] In order to overcome the inability of the existing liver model to maintain the structure and function of the liver for a long time in vitro, thereby performing functional tests such as drug metabolism and toxicity evaluation; the present invention provides a method such as figure 1 The preparation method of the tissue engineering liver model shown includes the following steps:

[0031] Step 1: Carry out the cultivation and expansion of seed cells;

[0032] Step 2: Prepare the liquid and matrix for model construction;

[0033] Step 3: Perform cell collection;

[0034] Step 4: Carry out model construction.

[0035] The first step: seed cell culture and expansion, including the following steps:

[0036] Step 1. Preparation of Matrigel: After freezing and thawing the Matrigel matrix overnight on ice at 4~8℃, use a pre-cooled pipette tip to mix the Matrigel matrix into a homogenate shape; use pre-cooled pure DMEM (according to 1:3 ~30 Dilute the Matrigel matrix; coat the diluted Matri...

Embodiment 2

[0047] Matrigel preparation: After freezing and thawing the Matrigel matrix overnight in a refrigerator at 4°C, mix it with a pre-cooled pipette tip to form a homogenate. Take a 50ml pre-cooled EP tube, add 28ml of pre-cooled pure DMEM medium (without serum, no additives and no antibiotics), add 2ml Matrigel matrix to dilute, all operations need to be performed on ice. Take out the culture vessel that needs to be coated, place it on ice, and coat the diluted Matrigel matrix in the desired culture vessel with the coating amount covering at least the entire growth surface of the vessel. Incubate for 1 hour at room temperature. Remove the unbound Matrigel matrix and rinse gently with pure DMEM medium.

[0048] Extraction and culture of primary hepatocytes: Weigh a certain amount of liver tissue, mechanically cut it, and digest it with 0.1% collagenase at 37°C for 30 minutes, and then digest with 0.125% pancreatin at 37°C for 30 minutes, 40 mesh Filter through a sieve to make a sin...

Embodiment 3

[0064] Matrigel preparation: After freezing and thawing the Matrigel matrix overnight in a refrigerator at 4°C, mix it with a pre-cooled pipette tip to form a homogenate. Take a 50ml pre-cooled EP tube, add 29ml of pre-cooled pure DMEM medium (without serum, no additives and no antibiotics), add 1ml Matrigel matrix to dilute, all operations need to be performed on ice. Take out the vessel to be coated, place it on ice, and coat the diluted Matrigel matrix in the required culture vessel, with the coating amount covering at least the entire growth surface of the vessel. Incubate for 1 hour at room temperature. Remove the unbound Matrigel matrix and rinse gently with pure DMEM medium.

[0065] Extraction and culture of primary hepatocytes: Weigh a certain amount of liver tissue, mechanically cut it, and digest it with 0.1% collagenase at 37°C for 30 minutes, and then digest with 0.125% pancreatin at 37°C for 30 minutes, 40 mesh Filter through a sieve to make a single cell suspensi...

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 relates to a preparation method of a tissue-engineered hepatic model. The preparation method of the tissue-engineered hepatic model comprises the following steps that step one, seed cells are cultivated and amplified; step two, liquid and matrix for model building are prepared; step three, cells are collected; and step four, the model is built. The tissue-engineered hepatic model iscomposed of primary hepatocytes, endothelial cells and hepatic stellate cells, a bile duct sample structure similar to natural hepatic tissue is formed, functions of albumin secretion and bile acid transport are achieved, cytochrome P450 activity is high, and the mitochondrial function is high. Compared with individually cultured hepatic cells, the albumin and urea expression level of the built hepatic tissue model is obviously improved, the tissue activity can be maintained in vitro for 28 days, long-term drug exposure can be realized, and a more sensitive and accurate test model is providedfor hepatotoxicity and efficacy evaluation based on vitro cells.

Description

Technical field [0001] The invention belongs to the technical field of regenerative medicine and tissue engineering, and specifically relates to a preparation method of a tissue engineering liver model. Background technique [0002] As the largest internal organ of the human body, the liver is responsible for many important physiological functions, such as bioconversion of sugar, metabolism of proteins, lipids, vitamins, hormones, synthesis and secretion of bile, etc. It is known as the human body’s "metabolic factory"; the liver is also the human body The "detoxification factory" is the most important organ in the human body where the toxicity of drugs and their metabolites manifests. Therefore, the liver has become an important target organ for drug toxicity and metabolism research. Hepatotoxicity is an important reason for the failure of new drug development. About two-thirds of the drugs entering Phase III clinical trials have to be terminated due to their liver toxicity. Dru...

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
Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071
CPCC12N5/067C12N2500/25C12N2500/36C12N2500/38C12N2501/11C12N2501/12C12N2501/237C12N2501/855C12N2502/14C12N2502/28C12N2533/50
Inventor 李润芝张勇杰金岩胡成虎卢永波
Owner GUANGDONG BOXI BIO TECH CO LTD
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com