Mouse liver organoid model, establishing method therefor and application of mouse liver organoid model

A technology for establishing methods and organoids, applied in the field of biomedicine, can solve the problems of inability to perform rapid high-throughput detection and analysis, limited practicability and long-term culture capacity, and limit the breadth and depth of research, so as to shorten the culture period and cost. Low, high cost effect

Active Publication Date: 2020-11-20
ACCURATE INT BIOTECHNOLOGY (GUANGZHOU) CO LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0002] Drug-induced hepatotoxicity is currently the most critical safety indicator for new drugs. Immortalized hepatic cell lines and animal models are still the "gold standard" for drug metabolism and toxicity testing, but their practicability and l

Method used

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  • Mouse liver organoid model, establishing method therefor and application of mouse liver organoid model
  • Mouse liver organoid model, establishing method therefor and application of mouse liver organoid model
  • Mouse liver organoid model, establishing method therefor and application of mouse liver organoid model

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Embodiment 1

[0046] This embodiment provides a method for establishing a mouse liver organoid model, comprising the following steps:

[0047] 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.

[0048] 2) Sample cutting: cut the tissue into small pieces of about 1 mm^3, and observe under the microscope whether there are cells leaking out.

[0049] 3) Tissue digestion: ① Collect the sheared tissue into a 15mL centrifuge tube with 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% double antibody for physiological Stop the digestion with saline, let the sediment stand still, suck off the supernatant as much as possible, and collect the cell pellet; ②Continue to add collagenase, dispase and DNaseI to the pellet, shake and digest at 37°C for 30 minutes, add 1% double-antibody normal saline...

Embodiment 2

[0056] This embodiment provides a method for establishing a mouse liver organoid model, and simultaneously uses the established liver organoid to investigate the efficacy of dexamethasone on liver cells, specifically including the following steps:

[0057] 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.

[0058] 2) Sample cutting: cut the tissue into small pieces of about 1 mm^3, and observe under the microscope whether there are cells leaking out.

[0059] 3) Tissue digestion: ① Collect the sheared tissue into a 15mL centrifuge tube with 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% double antibody for physiological Stop the digestion with saline, let the sediment stand still, suck off the supernatant as much as possible, and collect the cell pellet; ②Continue to ...

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Abstract

The invention provides a mouse liver organoid model, an establishing method therefor and application of the mouse liver organoid model. The establishing method comprises the following steps: withdrawing liver tissue of a mouse, and carrying out cleaning, shearing, digesting and filtering treatment sequentially, so as to obtain liver stem cells of the mouse; and subjecting the liver stem cells of the mouse to primary culture, propagation culture and differentiation culture sequentially, thereby obtaining the mouse liver organoid model. The liver organoid model obtained by employing the method is applied to pharmacologic, pharmacodynamic and toxicologic analysis. Compared with the conventional methods, the liver organoid model establishing method provided by the invention has the advantagesthat on one hand, the culture cycle is shortened greatly: the time cycle from material obtaining to model obtaining is about 14 to 20 days, the consumed time of the conventional culture method is 30 days, and thus, the feasibility of model application is improved obviously; and on the other hand, by the method provided by the invention, the success rate of mouse liver organoid culture is increasedobviously, and the growth speed of organoids of primary culture is accelerated.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to a mouse liver organoid model and its establishment method and application. Background technique [0002] Drug-induced hepatotoxicity is currently the most critical safety indicator for new drugs. Immortalized hepatic cell lines and animal models are still the "gold standard" for drug metabolism and toxicity testing, but their practicability and long-term culture capacity are very limited, which greatly affects The breadth and depth of research are limited. Liver toxicity experiments based on animal models are costly and have a long experiment cycle, making rapid high-throughput detection and analysis impossible. In vitro models capable of high-throughput analysis are urgently needed for drug efficacy and toxicity testing in hepatotoxicity studies and preclinical drug development. Mice are a commonly used model for drug safety evaluation. Establishing a mouse liv...

Claims

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Application Information

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IPC IPC(8): C12N5/074C12Q1/02
CPCC12N5/0672C12N2500/25C12N2500/32C12N2500/38C12N2501/11C12N2501/119C12N2501/155C12N2501/30C12N2501/345C12N2501/415C12N2501/999C12N2513/00G01N33/5088G01N33/94
Inventor 李刚杨志英陈泽新
Owner ACCURATE INT BIOTECHNOLOGY (GUANGZHOU) CO LTD
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