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Method for constructing liver and immune double-humanized large animal model by utilizing single stem cell transplantation

A stem cell differentiation and stem cell technology, applied in the fields of regenerative medicine and virology, experimental medicine, and clinical medicine, can solve the problems of no major breakthrough and great harm to human beings, and achieve the effect of high similarity with the human body

Pending Publication Date: 2021-10-22
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Infectious diseases such as hepatitis, AIDS, new coronavirus pneumonia, dengue fever, Ebola, Zika, malaria, sepsis, etc. have a wide range of epidemics, and the diseases caused by them are very harmful to humans. At present, there is no research on the diseases caused by these viruses big breakthrough

Method used

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  • Method for constructing liver and immune double-humanized large animal model by utilizing single stem cell transplantation
  • Method for constructing liver and immune double-humanized large animal model by utilizing single stem cell transplantation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Example 1: Transplantation of human bone marrow mesenchymal stem cells (hBMSCs) into miniature pigs with fulminant hepatic failure established a double humanized pig model of human bone marrow mesenchymal stem cells.

[0043] 1. Human bone marrow mesenchymal stem cells hBMSCs were cultured in DMEM medium containing 10% fetal bovine serum.

[0044] 2. Before jugular vein intubation, 8-10kg Chinese experimental minipigs were fasted for 12 hours and water was deprived for 8 hours. First, diazepam, ketamine and atropine were injected. After the animals were anesthetized, the drape was disinfected and intubated in the unilateral external jugular vein. After waking up, the animals were sent back to the breeding room. Modeling was usually performed 2 days after intubation.

[0045] 3. D-galactose (D-gal) at a dose of 1.5 g / kg was slowly injected into the animal through the jugular vein. During the whole experiment, all animals did not receive any treatment such as drugs.

[...

Embodiment 2

[0050] Example 2: Transplantation of human embryonic stem cell lines into miniature pigs with fulminant hepatic failure Establishment of a double humanized pig model of human embryonic stem cells.

[0051] 1. Culture human embryonic stem cells in DMEM medium containing 10% fetal bovine serum.

[0052] 2. Before jugular vein intubation, 8-10kg Chinese experimental minipigs were fasted for 12 hours and water was deprived for 8 hours. First, diazepam, ketamine and atropine were injected. After the animals were anesthetized, the drape was disinfected and intubated in the unilateral external jugular vein. After waking up, the animals were sent back to the breeding room. Modeling was usually performed 2 days after intubation.

[0053] 3. Acetaminophen (APAP) at a dose of 750 mg / kg was slowly injected into the animal through the jugular vein. During the whole experiment, all animals did not receive any treatment such as drugs.

[0054] 4. Put 1×10 7 Human embryonic stem cells were...

Embodiment 3

[0056] Example 3: Transplantation of human induced pluripotent stem cells (hiPSCs) into miniature pigs with fulminant hepatic failure Establishment of human induced pluripotent stem cell double humanized pig model.

[0057] 1. Introduce certain transcription factors into animal or human somatic cells through gene transfection technology, so that somatic cells can be directly restructured into pluripotent stem cells, and cultured in DMEM medium containing 10% fetal bovine serum.

[0058] 2. Before jugular vein intubation, 8-10kg Chinese experimental minipigs were fasted for 12 hours and water was deprived for 8 hours. First, diazepam, ketamine and atropine were injected. After the animals were anesthetized, the drape was disinfected and intubated in the unilateral external jugular vein. After waking up, the animals were sent back to the breeding room. Modeling was usually performed 2 days after intubation.

[0059] 3. D-galactose (D-gal) at a dose of 1.5 g / kg was slowly injecte...

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Abstract

The invention discloses a method for constructing a liver and immune double-humanized large animal model by utilizing single stem cell transplantation. The method comprises the following steps: acquiring human stem cells, transplanting a single stem cell into a large animal with liver injury, embedding human liver cells, forming a humanized immune system, and the like. The method can be used for researching the pathogenesis of human body diseases and researching, developing and screening new clinical medicines, and also can be used for obtaining an infectious disease model, a liver disease and drug metabolism model, a tumor growth and cancer immune model, an organ transplantation model, an autogenous immune disease model and the like which more accord with the development history of human diseases; and the humanized model which is convenient, simple, easy to obtain and high in similarity with a human body is provided for clinical liver disease treatment 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 method for constructing a liver and immune double humanized large animal model by transplanting a single stem cell. Background technique [0002] Experimental animal disease models are indispensable research tools for the study of the etiology and pathogenesis of human diseases, and the development of prevention techniques and drugs. However, the species differences between humans and animals, as well as the differences in the experimental results of disease models and clinical phenotypes, lead to serious consequences in the research and development of drugs on common animal models, such as a large number of toxic side effects, liver and kidney failure, etc. If human cells and tissues are introduced into the common animal model to establish a humanized animal model, it will provide an important tool for the ...

Claims

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Application Information

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IPC IPC(8): A01K67/027
CPCA01K67/0271A01K2207/12A01K2227/108A01K2267/0337A01K2267/0362A01K2267/0331A01K2267/0325A01K2267/03
Inventor 李君孙苏婉
Owner ZHEJIANG UNIV
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