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A method for evaluating drug hepatotoxicity based on liver organoid model

An evaluation method and organoid technology, applied in the field of biomedicine, can solve the problems of limited practicability and long-term culture ability, and achieve the effect of reducing the cost of new drug development, reducing adverse consequences, and easy operation

Active Publication Date: 2021-05-28
ACCURATE INT BIOTECHNOLOGY (GUANGZHOU) CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Hepatoma cell lines and animal models remain the "gold standard" for drug metabolism and toxicity testing, but their utility and long-term culture capabilities are limited

Method used

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  • A method for evaluating drug hepatotoxicity based on liver organoid model
  • A method for evaluating drug hepatotoxicity based on liver organoid model
  • A method for evaluating drug hepatotoxicity based on liver organoid model

Examples

Experimental program
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Embodiment 1

[0039] This embodiment provides a method for evaluating hepatotoxicity of chloroquine phosphate based on a liver organoid model, comprising the following steps:

[0040] 1. Establishment of mouse liver organoid model:

[0041] 1) Sample cleaning: the mouse liver tissue was washed several times with normal saline containing 1% double antibody, and the obvious fat tissue was removed.

[0042] 2) Sample cutting: Cut the tissue into small pieces of about 0.5mm^3.

[0043] 3) Tissue digestion: ①Disperse the sheared tissue in normal saline, centrifuge to remove the supernatant, add collagenase, dispase and DNaseI to the precipitate, shake and digest at 37°C for 30min, add 1% green chain double antibody The normal saline was used to stop the digestion, and the cell pellet was collected; ②Continue to add collagenase, dispase and DNaseI to the pellet, shake and digest at 37°C for 2 hours, add green chain normal saline containing 1% double antibody to stop the digestion, and let it sta...

Embodiment 2

[0052] This embodiment provides a method for evaluating the hepatotoxicity of mitoxantrone based on a liver organoid model, comprising the following steps:

[0053] 1. Establishment of mouse liver organoid model:

[0054] 1) Sample cleaning: the mouse liver tissue was washed several times with normal saline containing 1% double antibody, and the obvious fat tissue was removed.

[0055] 2) Sample cutting: Cut the tissue into small pieces of about 0.5mm^3.

[0056] 3) Tissue digestion: ①Disperse the sheared tissue in normal saline, centrifuge to remove the supernatant, add collagenase, dispase and DNaseI to the precipitate, shake and digest at 37°C for 30min, add 1% green chain double antibody The normal saline was used to stop the digestion, and the cell pellet was collected; ②Continue to add collagenase, dispase and DNaseI to the pellet, shake and digest at 37°C for 2 hours, add green chain normal saline containing 1% double antibody to stop the digestion, and let it stand P...

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Abstract

The present invention provides a method for evaluating drug hepatotoxicity based on a liver organoid model, comprising: step 1, establishment of a mouse liver organoid model: (1) taking mouse liver tissue, washing, shearing, digesting, and lysing in sequence and filtration treatment to obtain mouse liver stem cell precipitate; (2) the mouse liver stem cells are gelatinized and then cultured in culture medium to obtain a liver organoid model; step 2, the liver organoid model is treated with drugs: the drug to be tested is set Multiple concentration gradients were formed, and then the liver organoid model was given drug stimulation at different concentration gradients, and organoid samples were collected at different time points to observe their morphological changes and detect liver function indicators. The invention provides a more accurate in vitro method for detecting drug hepatotoxicity, which has short cycle and simple operation, can replace traditional cell line models and animal models, and is used for preclinical new drug development and clinical drug hepatotoxicity assessment.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to a drug hepatotoxicity evaluation method based on a liver organoid model. Background technique [0002] The liver's unique metabolic functions and relationship to the gastrointestinal tract make it an important target for drug and xenobiotic toxicity. Changes in liver metabolic activity from birth to adolescence with differential sensitivity to allotoxins. Hepatic drug metabolism, the often unbalanced production and detoxification process of toxic metabolites, can affect the degree of hepatotoxicity. In cases of severe toxicity, patients may develop hepatic failure. Drug-induced liver injury is currently the most common adverse event in the development of new drugs that leads to drug withdrawal due to safety, and liver toxicity discovered after approval for marketing also limits the use of many drugs. It would be a great boon to humans and experimental animals i...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/074C12Q1/02
CPCC12N5/0671C12N5/0672C12N2500/32C12N2500/38C12N2500/60C12N2501/11C12N2501/119C12N2501/237C12N2501/345C12N2501/727C12N2513/00G01N33/5014G01N33/5088
Inventor 李刚杨志英陈泽新
Owner ACCURATE INT BIOTECHNOLOGY (GUANGZHOU) CO LTD
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