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3D human liver organ model construction method, 3D human liver organ model and application of 3D human liver organ model

A construction method, 3D technology, applied in artificial cell constructs, 3D culture, biochemical equipment and methods, etc., can solve the problems of sensitivity improvement, achieve enhanced response sensitivity, eliminate screening differences, and have strong hepatotoxicity and damage effects Effect

Pending Publication Date: 2021-11-19
BEIJING DAXIANG BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The embodiment of the present disclosure provides a method for constructing a 3D human liver organ model, a 3D human liver organ model and its application, so as to solve the technical problem that the sensitivity of the existing 3D human liver organ model needs to be further improved in response to hepatotoxic drugs

Method used

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  • 3D human liver organ model construction method, 3D human liver organ model and application of 3D human liver organ model
  • 3D human liver organ model construction method, 3D human liver organ model and application of 3D human liver organ model
  • 3D human liver organ model construction method, 3D human liver organ model and application of 3D human liver organ model

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Embodiment 1

[0074] A method for constructing a 3D human liver organ model, comprising the following steps:

[0075] S11. Digest the 2D cultured human liver cancer cell line hepG2 in the growth phase with 0.25 (vt.)% trypsin into a single cell suspension, centrifuge and resuspend, and then prepare a density of 3.35-8.9×10 6 cell / mL single cell suspension;

[0076] S21. In a 1.5ml EP tube, add the single cell suspension and the matrix material according to the volume ratio of the single cell suspension and the matrix material at a ratio of 5.6:2.4, and pipette to mix evenly to obtain a mixed cell suspension ; Wherein, the matrix material is a mixed matrix material of collagen and matrigel, the volume ratio of collagen and matrigel is 1:1, and the concentration of collagen is 2.5mg / mL, and the concentration of matrigel is 5mg / mL.

[0077] S31. Using a pipette gun, quickly and high-throughput transfer the mixed cell suspension into the culture microwells of the 3D organ chip, then place it i...

Embodiment 2

[0088] A method for constructing a 3D human liver organ model, comprising the following steps:

[0089] S12. Resuscitate a primary human liver cell preserved in liquid nitrogen, dilute the primary human liver cell into a single cell suspension, centrifuge and resuspend, and then prepare a density of 4.45-44.5×10 6 cell / mL single cell suspension.

[0090] S22. In a 1.5ml EP tube, add the single cell suspension and the matrix material according to the volume ratio of the single cell suspension and the matrix material at a ratio of 5.6:2.4, and pipette to mix evenly to obtain a mixed cell suspension ; Wherein, the matrix material is a mixed matrix material of collagen and matrigel, the volume ratio of collagen and matrigel is 1:1, and the concentration of collagen is 2.5mg / mL, and the concentration of matrigel is 5mg / mL.

[0091] S32. Using a pipette gun, transfer and inoculate the mixed cell suspension into the culture microwells of the 3D organ chip in a rapid high-throughput ...

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Abstract

The invention relates to the technical field of biological tissue engineering, and discloses a 3D human liver organ model construction method which comprises the steps: preparing a single-cell suspension from human primary liver cells or mixed cells of the human primary liver cells and non-parenchymal liver cells, or a human liver cancer cell line, and mixing the single-cell suspension with a matrix material to obtain a mixed cell suspension; inoculating the mixed cell suspension into culture micropores of a 3D organ chip, and culturing at the temperature of 37 DEG C to obtain a gel-forming 3D organ chip; and adding a culture medium into the liquid storage hole, and culturing to obtain the 3D human liver organ model. Compared with other 2D human liver organ models, the constructed 3D human liver organ model has the advantages that the response sensitivity of the constructed 3D human liver organ model to hepatotoxic drugs is obviously enhanced, and the hepatotoxic injury effect of the reported hepatotoxic drugs is stronger. Compared with an animal model, the 3D human liver organ model can effectively eliminate screening differences caused by species differences. Therefore, the 3D human liver organ model can detect more drugs with hepatotoxic injury in vitro, and the drugs are closer to clinical reports.

Description

technical field [0001] This application relates to the technical field of biological tissue engineering, for example, to a method for constructing a 3D human liver organ model, a 3D human liver organ model and applications thereof. Background technique [0002] Currently, drug-induced liver injury is the main reason for drug clinical attrition, precautionary warnings, and post-market withdrawal. Therefore, more predictive methods are needed to assess the risk of hepatotoxicity in the drug discovery process. Usually the US FDA and the European Medicines Agency require candidate new drugs to be evaluated for toxicity on rodent (such as mice) and non-rodent (such as dogs) animals before clinical trials. Available data show that the effective predictive rate of toxicity in rats and dogs is only 71% of that in humans. Through the investigation and comparison of target organ toxicity, it was found that the consistent rate of hepatotoxicity in animal and human clinical phase I is...

Claims

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Application Information

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IPC IPC(8): C12N5/09C12N5/071C12N5/02C12Q1/02
CPCC12N5/0693C12N5/0671G01N33/5076C12N2503/02C12N2513/00G01N2500/10A61L27/26A61L27/38G01N33/5014G01N33/5088C12N5/0068C12N5/0697C12N2509/00G01N33/5082
Inventor 肖荣荣周宇
Owner BEIJING DAXIANG BIOTECH CO LTD