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Method for constructing humanized chronic hepatitis B mouse model by means of stem cell

A stem cell and mouse model technology, applied in the fields of clinical medicine, regenerative medicine and virology, and experimental medicine, can solve problems such as difficulty in establishing research models, long experimental periods, and limiting the optimization of treatment plans.

Active Publication Date: 2017-09-15
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Hepatotropic virus can only cause disease in higher primates. Animal experimental models often need to use apes. The cost of ape experiments is high, the operation is complicated, and the experimental cycle is long. Therefore, it is necessary to establish Small animal models have great scientific significance and application value
However, since small animals cannot be infected with hepatotropic virus, it is difficult to establish a small animal research model of hepadnavirus, which seriously hinders the research on the mechanism, progress and outcome of viral hepatitis, and largely limits the optimization of treatment options.

Method used

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  • Method for constructing humanized chronic hepatitis B mouse model by means of stem cell
  • Method for constructing humanized chronic hepatitis B mouse model by means of stem cell
  • Method for constructing humanized chronic hepatitis B mouse model by means of stem cell

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Example 1: figure 1 Schematic diagram for the construction of a humanized FRGS mouse model of bone marrow mesenchymal stem cells. Human bone marrow mesenchymal stem cells (hBMSCs) were transplanted into FRGS mice with fulminant liver failure to establish a humanized FRGS mouse model of bone marrow mesenchymal stem cells.

[0046] 1. Human bone marrow mesenchymal stem cells hBMSCs were cultured in DMEM medium containing 10% fetal bovine serum.

[0047] 2. FRGS mice gradually reduced the drug 2-(2-nitro-4-trifluoromethylbenzyl)-cyclohexane-1,3-dione (NTBC) and injected 0.2mg / kg anti-Fas antibody (JO2), to establish a mouse model of liver failure.

[0048] 3. Inject 500ul 1×10 through the portal vein 6 hBMSCs.

[0049] 4. Continuous JO2 injection at 2, 5, and 8 days after transplantation.

[0050] 5. Inject each mouse with type A, B, C, and D 1*10 through the tail vein 6 Hepatitis B virus.

[0051] 6. After 1 week, 2 weeks, and 4 weeks after the injection of hepatiti...

Embodiment 2

[0053] Example 2: figure 2 Schematic diagram for the construction of an embryonic stem cell humanized uPA mouse model. Human embryonic stem cell lines were transplanted into homozygous uPA mice to establish an embryonic stem cell humanized uPA mouse model.

[0054] 1. Culture human embryonic stem cells in DMEM medium containing 10% fetal bovine serum.

[0055] 2. Obtain a homozygous uPA / SCID mouse model.

[0056] 3. Inject 500ul 1×10 via the spleen at 8 weeks of birth 6 Human embryonic stem cells.

[0057] 4. Inject 1*10 per mouse via the tail vein 7 Hepatitis C virus.

[0058] 5. After 1 week, 2 weeks, and 4 weeks after the injection of hepatitis C virus, the viral load and liver function status were detected every 4 weeks to confirm the successful establishment of the model.

[0059] figure 2 Using uPA mice, uPA will spontaneously form liver damage, and then use human embryonic stem cell transplantation to differentiate into liver cells, and finally inject hepatitis C ...

Embodiment 3

[0060] Example 3, image 3 Schematic diagram for the establishment of a normal mouse model of induced pluripotent stem cell humanized galactosamine. Human induced pluripotent stem cells (hiPSCs) were injected into normal immune fulminant liver failure mice to establish a normal mouse model of induced pluripotent stem cell humanization.

[0061] 1. Introduce certain transcription factors into animal or human somatic cells through gene transfection technology, so that the somatic cells can be directly restructured into pluripotent stem cells, and cultured in DMEM medium containing 10% fetal bovine serum.

[0062] 2. Each mouse was intraperitoneally injected with 1.5 g / kg of galactosamine to establish a mouse model of liver failure.

[0063] 3. Inject 500ul 1×10 through the liver 6 Human induced pluripotent stem cells.

[0064] 4. Inject 1*10 per mouse via the tail vein 5 Hepatitis E virus.

[0065] 5. After 1 week, 2 weeks, and 4 weeks after the injection of hepatitis E viru...

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Abstract

The invention discloses a method for constructing a humanized chronic hepatitis B mouse model by means of a stem cell. The method comprises the following steps: obtaining a human stem cell, transplanting the stem cell into a mouse with liver damage, infecting a humanized mouse with HBV, etc. It is discovered that by inducing severe liver damage and transplanting the human stem cell, human-derived liver cells in the liver of the mouse have a high chimerism of 50-95%, and humanized immunological cells can be continuously separated in the spleen, blood, liver, marrow and other organs of the mouse, thus forming a mouse model with humanized liver and immunological cells; and the humanized mouse is infected with various hepatotropic viruses, thus forming a hepatotropic virus infected humanized mouse model. The technology for constructing a humanized mouse model is used for constructing a model for researching hepatotropic virus infection. Besides, the idea in the technical scheme can be also used for constructing other humanized organ models; and the technical scheme provides a convenient, simple and accessible humanized model for clinical liver disease treatment and research.

Description

technical field [0001] The invention belongs to the fields of clinical medicine, experimental medicine, regenerative medicine and virology, and specifically relates to a new method for constructing a humanized mouse model and being used for the research of viral hepatitis and other human diseases. Background technique [0002] Hepatotropic virus (hepatitis A virus, hepatitis B virus, hepatitis C virus, etc.) has a wide range of epidemics, and millions of people die every year from liver failure, cirrhosis and hepatocellular carcinoma caused by viral hepatitis. Hepatotropic virus can only cause disease in higher primates. Animal experiment models often need to use apes. The cost of ape experiments is high, the operation is complicated, and the experiment period is long. Therefore, it is necessary to establish Small animal models have great scientific significance and application value. However, because small animals cannot be infected with hepatotropic virus, it is difficult...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01K67/027
CPCA01K67/0271A01K2267/0337A01K2207/20A01K2207/12A01K2227/106C12N5/0667C12N5/0663C12N5/0696C12N5/0606A01K2227/105A01K67/0278A61K9/0053A61K9/0019
Inventor 李君李江孙苏婉李兰娟
Owner ZHEJIANG UNIV
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