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Artificial liver structure containing bile duct and liver tissue as well as preparation method and application of artificial liver structure

A technology of liver tissue and structure, applied in the fields of biomaterials and biomedical engineering, can solve the problems of blank and unseen in the technical field of artificial liver structure, and achieve the effect of great difficulty in culturing

Active Publication Date: 2022-04-22
TSINGHUA UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the technical field of constructing artificial liver structures containing bile duct tissue and liver tissue is still blank, and no relevant reports have been seen

Method used

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  • Artificial liver structure containing bile duct and liver tissue as well as preparation method and application of artificial liver structure
  • Artificial liver structure containing bile duct and liver tissue as well as preparation method and application of artificial liver structure
  • Artificial liver structure containing bile duct and liver tissue as well as preparation method and application of artificial liver structure

Examples

Experimental program
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preparation example Construction

[0072] The present invention also provides a method for preparing the above-mentioned artificial liver structure containing bile duct tissue and liver tissue, comprising the following steps:

[0073] (1) Uniformly mixing the biocompatible material with cells to obtain a precursor solution containing cells;

[0074] (2) preparing the precursor solution into a three-dimensional hydrogel structure according to a pre-designed structure;

[0075] (3) Post-processing of the three-dimensional hydrogel structure;

[0076] (4) In vitro culture and / or cell differentiation of the three-dimensional hydrogel structure to obtain the artificial liver tissue (artificial liver structure) containing bile duct tissue and liver tissue.

[0077] Further, the artificial liver tissue is applied to fields such as drug detection, environmental monitoring, toxicological detection, tissue engineering, regenerative medicine, new drug research and development, liver tissue development, and disease occurr...

Embodiment 1

[0096] Example 1 Preparation of artificial liver structure using dual-nozzle bio-3D printing technology

[0097] This embodiment provides an artificial liver structure that can be constructed by bio-3D printing equipment, such as figure 2 shown.

[0098] 1. Planar culture of hepatocytes

[0099] Human normal hepatocytes (L-02) were purchased from Procell, the product number is HL-7702. Human normal hepatocytes were cultured on a flat surface using the expansion medium, and the medium was replaced every 2 days. When the cells reached 85% confluency, they were subcultured at a ratio of 1:3. The composition of the culture medium of human normal hepatocytes is: Williams' Medium E medium (Sigma, W4125) supplemented with 10% FBS serum (Gibco, 16000), 1% penicillin and streptomycin (Gibco, 15140122), 20ng / mL liver Cell growth factor (HGF, R&D Systems) and 50 μM β-mercaptoethanol.

[0100] 2. Planar culture of bile duct epithelial cells

[0101] Human intrahepatic bile duct epit...

Embodiment 2

[0141] Example 2 Preparation of Artificial Liver Structural Body by Casting Method

[0142] This embodiment provides a method for preparing an artificial liver structure by casting, such as Figure 5 with Image 6 shown. Including the following steps:

[0143] 1. Induction of liver stem cells

[0144] Induced pluripotent stem cells (iPSCs) were used to induce differentiation to obtain liver stem cells. The specific implementation steps are as follows: iPS cells (Anhui Zhongsheng Suyuan Biotechnology Co., Ltd.) were dissociated using cell dispase (Dispase, Roche Diagnostics) and seeded on BD Matrigel Growth Factor Reduced Basement MembraneMatrix (Becton Dickinson). Subsequently, iPSCs were cultured in L-WNT3A(CRL2647; ATCC)-expressing cell-conditioned RPMI 1640 medium (Sigma) for 4 days to differentiate to the defined endoderm stage. The medium components included: 100ng / mL Activin A (R&D Systems), 1% GlutaMAX (Thermo Fisher Scientific), 1% penicillin streptomycin (Gibco),...

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Abstract

The invention provides an artificial liver structure containing bile duct and liver tissue as well as a preparation method and application of the artificial liver structure. The artificial structure contains various cells such as parenchymal hepatic cells and bile duct epithelial cells. The hepatocytes are in a liver plate shape or a cluster shape, and the bile duct epithelial cells are in a bile duct-like three-dimensional vascular network shape. The artificial liver structure has an albumin secretion function and a nitrogen metabolism function of a mature liver tissue, and expresses a symbolic protein marker of a mature liver cell. The artificial liver structural body also has a bile duct network structure formed by connecting bile duct epithelial cells and expresses a bile duct epithelial cell marker. According to the preparation method of the artificial structure, cells from various sources can be adopted, and customized design and batch production can be carried out according to needs. The artificial liver structure provided by the invention is a liver model with wide application, and is used in the fields of preclinical drug detection, environmental monitoring, toxicology detection, tissue engineering, regenerative medicine, new drug research and development, research on liver tissue development, research on disease occurrence and development and the like.

Description

technical field [0001] The invention relates to the fields of biomaterials and biomedical engineering, in particular to an artificial liver structure containing bile duct and liver tissue, its preparation method and application. Background technique [0002] The liver is the most important detoxification organ in the human body. Almost all exogenous and endogenous toxic substances must be decomposed by the liver to be excreted. The main cause of adverse drug reactions is drug-induced liver injury. Nearly 90% of compounds that pass preclinical drug screening experiments ultimately fail clinical trials, with one-third of compound failures attributed to their drug toxicity. There are also frequent recalls of drugs already on the market, about 50% of which are due to drug-induced liver injury. Due to the ethical and safety constraints of human trials, the commonly used drug testing models at this stage are animal models and planar cultured cells, but the results are often diffi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61L27/38A61L27/52A61L27/50C12N5/071
CPCA61L27/3804A61L27/3813A61L27/3834A61L27/52A61L27/50C12N5/0697A61L2430/28C12N2513/00C12N2502/09C12N2502/14C12N2503/02C12N2533/56C12N2533/40C12N2501/155C12N2501/18C12N2501/117C12N2501/11C12N2501/237C12N2500/30C12N2501/16C12N2500/62
Inventor 姚睿徐铭恩冯璐梁少君刘明
Owner TSINGHUA UNIV
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