Construction method and application of in-vitro three-dimensional human liver tissue

A construction method and technology of liver tissue, applied in artificial cell constructs, tissue screening, tissue regeneration, etc., can solve the problems of uncontrollable size and size, poor functional characteristics, etc., and achieve the effect of fast printing speed and high efficiency

Inactive Publication Date: 2017-07-04
TSINGHUA UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0008] In view of the disadvantages of poor functional characteristics and uncontrollable size of the in vitro human liver tissue constructed in the pri...

Method used

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  • Construction method and application of in-vitro three-dimensional human liver tissue
  • Construction method and application of in-vitro three-dimensional human liver tissue
  • Construction method and application of in-vitro three-dimensional human liver tissue

Examples

Experimental program
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Effect test

Embodiment 1

[0048] This embodiment relates to a method for constructing a three-dimensional human liver tissue in vitro, comprising the following steps:

[0049] Preparation steps:

[0050] 1) Preparation of bioink original solution

[0051] Gelatin solution: mix gelatin powder (Sigma, G1890) with deionized water at a mass ratio of 15:100, heat at 80°C for 3 hours to dissolve it evenly, then divide and store at 4°C or -20°C Store at low temperature. Incubate in the cell culture incubator for 10 minutes before each use to make it melt into a homogeneous solution.

[0052] Sodium alginate solution: mix sodium alginate powder (Sigma, A0682) with deionized water at a mass ratio of 4:100, heat at 80°C for 3 hours to dissolve it evenly, then pack it, and store it at 4°C Or store at -20°C. Incubate in the cell culture incubator for 10 minutes before each use to make it melt into a homogeneous solution.

[0053] 2) Cultivation of seed cells

[0054] HepaRG cells (sigma, HPRGC1) were culture...

Embodiment 2

[0061] Compared with Example 1, the difference of this example is that in step 4), the cell induction culture medium is 2% DMSO added to the cell expansion culture medium.

Embodiment 3

[0063] Compared with Example 1, the difference of this example is that in step 4), the cell induction culture medium is 1% DMSO added to the cell expansion culture medium.

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Abstract

The invention relates to a construction method for in-vitro three-dimensional human liver tissue. The construction method comprises the following steps: (1) uniformly mixing a solution of seed cells with an aqueous solution of biological ink so as to obtain a cell printing solution; (2) under the control of a computer, uniformly extruding the cell printing solution with a cell printer at a set speed according to a designed path and allowing the cell printing solution to rapidly coagulate at a certain temperature so as to form gel microfilaments, and repeating the above operations so as to finish construction of a multilayer cell structure in a layer-upon-layer scanning manner; (3) subjecting the multilayer cell structure to secondary crosslinking and culturing the multilayer cell structure with a cell culture solution so as to obtain a stable multilayer cell structure; and (4) culturing the stable multilayer cell structure with an induction medium so as to obtain the in-vitro three-dimensional human liver tissue. The method provided by the invention can construct in-vitro three-dimensional human liver tissue with good functionality, and the constructed in-vitro three-dimensional human liver tissue is applicable to fields like treatment of hepatopathy, medicinal development, medicine screening, etc.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a three-dimensional bionic human liver tissue construction method in vitro. Background technique [0002] The liver is the largest internal organ in the human body and is responsible for many important functions such as detoxification, metabolism, bile secretion and immune defense. my country is a big country with liver disease. About 300,000 people die of liver disease every year, and about one tenth of the population is a carrier of hepatitis B virus. The cost of liver disease treatment exceeds 30 billion. For severe diseases such as liver cancer and acute liver failure, the best way of treatment is liver transplantation, but the limitation of organ donor sources is still an unavoidable problem. On the other hand, the average R&D cost of a new drug is about US$5 billion. After 12 years of dedicated investment, most of the funds and time are spent on compound screening and drug tes...

Claims

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Application Information

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IPC IPC(8): C12N5/074C12N5/071A61L27/26A61L27/38C12Q1/02
CPCA61L27/26A61L27/3804A61L27/3839A61L2430/28C12N5/0602C12N5/0645C12N5/0656C12N5/0671C12N5/0672C12N2503/02C12N2503/04C12N2513/00C12N2537/10C12N2539/00G01N33/5008G01N33/5014G01N33/5082C08L5/04C08L5/08
Inventor 姚睿孙伟
Owner TSINGHUA UNIV
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