Liver tissue-like structural body as well as preparation method and application thereof

A liver tissue and structure technology, applied in the fields of biomaterials and biomedical engineering, can solve the problems of immature physiological functions of cells, time-consuming differentiation process, and inability to obtain liver cells, etc., so as to facilitate large-scale research and application, and facilitate observation , the effect of high yield

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

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Problems solved by technology

However, stem cell differentiation research at this stage is facing a common problem: uncontrollable differentiation (differentiation is not uniform, low efficiency and poor reproducibility), the obtained cells are immature and have poor physiological functions, the differentiation process takes a long time and the yield is low, etc. , it is impossible to obtain a large number of stem cell-derived hepatocytes with no intra-batch variation and stable functions, which cannot meet the needs of basic and applied research on tissue development, liver regeneration, and the occurrence and development of liver diseases

Method used

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  • Liver tissue-like structural body as well as preparation method and application thereof
  • Liver tissue-like structural body as well as preparation method and application thereof
  • Liver tissue-like structural body as well as preparation method and application thereof

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preparation example Construction

[0075] The present invention also provides a method for preparing the aforementioned artificial liver tissue with excellent physiological functions, comprising the following steps:

[0076] (1) Uniformly mixing the biocompatible material with cells to obtain a precursor solution containing cells;

[0077] (2) preparing the precursor solution into a three-dimensional hydrogel structure according to a pre-designed structure;

[0078] (3) post-processing the three-dimensional hydrogel structure;

[0079] (4) In vitro culture and / or cell-induced differentiation of the three-dimensional hydrogel structure to obtain bionic artificial liver tissue;

[0080]Further, the artificial liver tissue is applied in the fields of liver tissue development research, liver tissue regeneration research, liver disease occurrence and development research, preclinical drug testing, and new drug testing and development.

[0081] According to the method above, an artificial liver tissue with excellen...

Embodiment 1

[0099] Example 1 Using 3D bioprinting to prepare artificial liver tissue with excellent physiological functions

[0100] This embodiment provides an artificial liver tissue with excellent physiological functions constructed by bio-3D printing equipment, such as figure 1 shown.

[0101] 1. Acquisition and culture of liver progenitor cells and human hepatocytes

[0102] Acquisition and culture of liver progenitor cells: Induced pluripotent stem cells were used to induce differentiation to obtain liver progenitor cells. The specific implementation steps are: inoculating induced pluripotent stem cells on Matrigel substrate (Becton Dickinson) and culturing for 3-4 days. Then cultured for 4 days to differentiate the induced pluripotent stem cells to the defined endoderm stage. The composition of the culture medium was: L-WNT3A (CRL2647; ATCC)-expressing cell-conditioned RPMI 1640 culture medium (Life, C11875500BT), 100ng / ml Activin A (ActivinA, Gibco, PHG9014), 1% GlutaMAX TM ...

Embodiment 2

[0128] Example 2 Preparation of Artificial Liver Tissue with Excellent Physiological Functions by Casting Method

[0129] This embodiment provides artificial liver tissues with various structures and excellent physiological functions prepared by the casting method, including the following steps:

[0130] 1. Liver stem cell culture

[0131] HepaRG is a liver stem cell obtained from human liver cancer tissue, which has the ability to bidirectionally differentiate into liver cells and bile duct epithelial cells. HepaRG cells (Sigma, HPRGC1) were cultured in cell expansion medium, and the composition of cell expansion medium was: Williams' Medium E medium (Sigma, W4125) was added with 10% FBS serum (Gibco, 16000), 0.05% insulin (Sigma, I9278), 5×10 -5 M hydrocortisone succinate (Sigma, H4881), 1% penicillin streptomycin (Gibco, 15140122) and 1% GlutaMAX TM (Gibco, 35050061). When the cells were 90% confluent, they were subcultured at a ratio of 1:5, and the culture medium was ...

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Abstract

The invention provides a liver-like tissue structure as well as a preparation method and application thereof. The cell phenotypes in the liver tissue-like structural body are highly consistent, and are uniformly dispersed in the whole structure in the form of small-size cell clusters. The liver tissue has a highly bionic physiological function, not only positively expresses symbolic genes and proteins of mature liver tissues, but also has higher physiological functions of albumin secretion, nitrogen metabolism, urea synthesis, detoxification, drug metabolism and the like. According to the preparation method of the liver tissue-like structural body, cells from various sources can be adopted, and customized design and batch production can be realized according to requirements. The liver tissue-like structural body disclosed by the invention is a liver model with wide application, and is used in the fields of liver tissue development research, liver tissue regeneration research, liver disease occurrence and development research, preclinical drug detection, new drug test and development, drug toxicology research and the like.

Description

technical field [0001] The invention relates to the fields of biomaterials and biomedical engineering, in particular to a liver tissue-like structure and its preparation method and application. Background technique [0002] The liver is the largest solid organ in the human body, accounting for approximately 2% of the total body weight. As the largest digestive gland and metabolic center in the human body, the liver plays an important role in maintaining the metabolic homeostasis of the human body. The liver is responsible for the synthesis of nutrients and is responsible for the synthesis and secretion of serum proteins, amino acids and glycogen in the human body. The liver also undertakes the detoxification function, metabolizing metabolic wastes, drugs and other compounds from endogenous and exogenous sources. In addition, the liver is also responsible for the metabolism and storage of substances including amino acids, vitamins, lipids and carbohydrates. [0003] Due to...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071
CPCC12N5/0671C12N2513/00C12N2533/54C12N2533/56C12N2533/40C12N2501/155C12N2501/115C12N2501/12C12N2501/998C12N2500/38C12N2501/16
Inventor 姚睿冯璐刘明梁少君
Owner TSINGHUA UNIV
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