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Construction method and application of rodent model of novel coronavirus infection

An animal model, coronavirus technology, applied in animal husbandry, etc., can solve the problem that the new coronavirus model cannot normally express the human ACE2 receptor protein lung physiological state, and achieve easy operation, low construction cost, and high infection efficiency Effect

Active Publication Date: 2021-11-30
TONGJI UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] Therefore, the technical problem to be solved by the present invention is to overcome the defects that the new coronavirus model constructed in the prior art cannot normally express human ACE2 receptor protein and simulate the physiological state of normal human lungs, thereby providing a new coronavirus SARS- Rodent animal model of CoV-2 infection and its construction method and use

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  • Construction method and application of rodent model of novel coronavirus infection
  • Construction method and application of rodent model of novel coronavirus infection
  • Construction method and application of rodent model of novel coronavirus infection

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Embodiment 1

[0049] This embodiment provides a method for constructing a rodent model of novel coronavirus SARS-CoV-2 infection using a bleomycin injury model and Tdtomato fluorescent protein-labeled human bronchial basal cells, comprising the following steps:

[0050] (1) Labeling of human bronchial basal cells: Mix lentiviral packaging plasmids pHIV-dTomato, psPAX2 and pMD2.G at a mass ratio of 5:3.75:1.25, transfect packaging cells 293T, and co-culture for 72 hours , collect the culture supernatant containing Tdtomato lentivirus, and centrifuge at 2000g for 10 minutes at 4°C to remove cell debris. The supernatant was filtered through a 0.45 μm filter and collected in an ultracentrifuge tube. Concentrate by ultracentrifugation at 25,000 g at 4°C to obtain a Tdtomato lentivirus suspension. After determining the titer of Tdtomato lentivirus, according to every 1×10 6 Individual bronchial basal cells added 10 7 The Tdtomato lentivirus of TU was co-cultured, and the medium was changed aft...

Embodiment 2

[0064] The present embodiment provides a kind of method adopting bleomycin injury model and human bronchial basal layer cell to construct the rodent model infected by novel coronavirus SARS-CoV-2, comprising the following steps:

[0065] (1) Construction of lung injury model: prepare 6-8 weeks old NOD.CB17-Prkdc scid / NcrCrl mice were divided into no-transplant control group, 4-day post-infection group and 21-day post-infection group, with one mouse in each group. After inhalational anesthesia with isoflurane, the trachea was intubated, and the bleomycin-PBS solution was administered to the lungs of the three groups of mice through the catheter through a syringe, with a volume of 50 μL for each mouse. The dose is 50 μg bleomycin / monkey.

[0066] (2) Cell transplantation: 7 days after administration of bleomycin, the mice in the 4-day post-infection group and the 21-day post-infection group were anesthetized with isoflurane again and then intubated. Take the human bronchial b...

Embodiment 3

[0070] This embodiment provides a method for constructing a rodent model of novel coronavirus SARS-CoV-2 infection using lipopolysaccharide combined with porcine trypsin-induced lung injury model and human bronchial basal layer cells, comprising the following steps:

[0071] (1) Construction of lung injury model: prepare three 6-8 week old NOD.CB17-Prkdc scid / NcrCrl mice. After anesthesia with isoflurane, each mouse was given a premixed solution of 40 μL porcine trypsin (10 U / mL) and 0.8 μL lipopolysaccharide (10 mg / mL) through the glottis into the trachea (the solvent was sterile PBS) . All mice were injured by continuous instillation for 3 days, once a day, and the volume of each instillation premix was 40.8 μL, and the lung injury model was established.

[0072] (2) Cell transplantation: Cell transplantation was carried out on the second day after the establishment of the lung injury model. After anesthesia with isoflurane, the human bronchial basal layer cell suspension...

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Abstract

The present invention relates to the technical field of animal model construction, and specifically provides a rodent animal model infected by a novel coronavirus SARS-CoV-2 and its construction method and application. The construction method includes the following steps: construction of a lung injury animal model: Immunodeficient animals were selected as objects to construct a lung injury animal model; cell transplantation: transplanting human-derived lung epithelial cells into the lungs of the lung injury animal model; SARS-CoV-2 virus infection: transplanting SARS-CoV-2 virus Infect the lungs of animals after the cell transplantation step to establish a rodent model infected by the new coronavirus SARS‑CoV‑2. This method is not only simple in construction process, easy to operate, low in construction cost, and can simulate Normal human lung tissue structure and expression of human ACE2 receptor protein.

Description

technical field [0001] The invention relates to the technical field of animal model construction, in particular to a rodent animal model infected by a novel coronavirus SARS-CoV-2 and its construction method and application. Background technique [0002] Novel coronavirus disease (coronavirus disease 2019, COVID-19) is a new infectious disease, a respiratory disease caused by severe acute respiratory syndrome coronavirus 2 (severe acute respiratory syndrome coronavirus 2, SARS-CoV-2) , broke out on a global scale, and its high infection rate and death rate threaten the whole world. After SARS-CoV-2 infects the human body, the most common symptoms are cough, fever, shortness of breath, and dyspnea. Severe infection may lead to severe acute respiratory syndrome, renal failure, and even death. Studies have found that SARS, MERS, SARS-CoV-2 and other coronavirus infections can cause an imbalance in the body's immune regulatory network, trigger cytokine storm syndrome (cytokines...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01K67/027
Inventor 左为张婷
Owner TONGJI UNIV
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