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Construction method of three-dimensional liver microtissue unit for bio-artificial liver support system

A liver support system and bioartificial technology, applied in the field of construction of three-dimensional micro-liver tissue units, can solve the problems that it is difficult to play a supporting and regulating role, and it is difficult to form high-function three-dimensional micro-liver tissues. Conducive to clinical application, improve the effect of activity and function

Inactive Publication Date: 2016-11-23
DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, there are still two key problems in the existing three-dimensional hepatocyte culture system
First of all, most of the existing systems only use one type of stromal cells to support liver cells, which cannot well simulate the coexistence of multiple important cells in liver tissue in vivo, and it is difficult to form a high-function three-dimensional micro-liver tissue
In addition, the existing system mainly improves the biological function of the scaffold by adding ingredients such as collagen and hyaluronic acid. However, these ingredients are far from the actual ingredients of the liver extracellular matrix in vivo, and it is difficult to play a sufficient supporting and regulating role.

Method used

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  • Construction method of three-dimensional liver microtissue unit for bio-artificial liver support system
  • Construction method of three-dimensional liver microtissue unit for bio-artificial liver support system
  • Construction method of three-dimensional liver microtissue unit for bio-artificial liver support system

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Effect test

Embodiment 1

[0034] Example 1: The effect of multi-cell co-culture on the function of three-dimensional micro-liver tissue

[0035] Anesthetize, depilate, and disinfect the miniature pigs, make a cross incision in the middle of the abdomen, infuse the hepatic artery with heparin solution for anticoagulation, intubate the portal vein and fix it, and perfuse it with 0.25% trypsin and 0.02% EDTA at 37°C with a peristaltic pump at a rate of 5 mL / min Perfuse the liver with about 2L of PBS solution until the liver is yellowish brown, cut off the perihepatic blood vessels, take the complete left outer lobe of the liver and store it at -80°C for 24 hours, cut the liver lobe into 3mm thick 2cm×2cm tissue slices with The liver tissue piece and the decellularized solution were mixed in a ratio of 1:15 (w / v) in a bottle and placed in a constant temperature shaker at 200 rpm at 4°C. According to double distilled water for 24 hours, 2% (v / v) Triton X-100 and 0.1% (v / v) ammonia water for 96 hours, 0.1% (...

Embodiment 2

[0038] Example 2 Effect of acellular matrix on the function of three-dimensional micro-liver tissue

[0039] Anesthetize, depilate, and disinfect the minipigs, make a cross incision in the middle of the abdomen, infuse the hepatic artery with heparin solution for anticoagulation, insert and fix the portal vein, and perfuse it with a peristaltic pump at a rate of 5 mL / min at 37°C containing 0.25% trypsin and 0.02% EDTA Perfuse the liver with about 2 L of PBS solution until the liver is yellowish brown, cut off the perihepatic blood vessels, take the complete left outer lobe of the liver and store it at -80°C for 24 hours, cut the liver lobe into 3mm thick 2cm×2cm tissue slices with The liver tissue piece and the decellularized solution were mixed in a ratio of 1:15 (w / v) in a bottle and placed in a constant temperature shaker at 200 rpm at 4°C. According to double distilled water for 24 hours, 2% (v / v) Triton X-100 and 0.1% (v / v) ammonia water for 96 hours, 0.1% (w / v) SDS for 2...

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Abstract

The invention discloses a construction method of a three-dimensional liver microtissue unit for a bio-artificial liver support system. The construction method is characterized by comprising the steps that decellularized scaffold components of a pig liver are encapsulated by microcapsules prepared from sodium alginate-polycations, liver cells, vascular endothelial cells and fibroblasts are composited, and the functional three-dimensional liver microtissue unit is constructed through cell self-assembly and applied to the bio-artificial liver support system. According to the method, operation is easy, cost is low, the liver cells, the vascular endothelial cells and the fibroblasts are self-assembled into three-dimensional liver microtissue by simulating interaction between the cells and an extracellular matrix, interaction between the cells and soluble factors and interaction between the cells and the cells in a liver tissue microenvironment, the three-dimensional liver microtissue has good synthesis, secretion and detoxification functions, and therefore the three-dimensional liver microtissue can serve as the microunit to be applied to the bio-artificial liver support system.

Description

technical field [0001] The invention relates to the field of bioartificial liver support systems, in particular to a method for constructing three-dimensional micro-liver tissue units used in bioartificial liver support systems. Background technique [0002] The liver is the most important metabolic organ in the human body. The bioartificial liver (BAL) support system can better replace the functions of liver synthesis, metabolism and secretion. application prospects. However, hepatocytes under conventional two-dimensional culture conditions quickly lose their main functions such as synthesis and detoxification, so how to obtain high-functioning hepatocytes is one of the core problems faced by the application of bioartificial livers. [0003] Studies have shown that the function of liver cells is closely regulated by the microenvironment of the liver tissue, and a variety of cells coexist in the microenvironment of the liver tissue. There are important interactions between...

Claims

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Application Information

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IPC IPC(8): C12N5/071C12N11/10C12N11/08C12N11/04
Inventor 马小军孙东升刘洋孙广炜王淑君
Owner DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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