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Breast cancer bone metastasis mouse model molding method

A mouse model, breast cancer technology, applied in the fields of botanical equipment and methods, biochemical equipment and methods, retro RNA viruses, etc., can solve the problem that breast cancer cells are not easy to transfer to other parts, the transfer of metastasis is less, and it is inconvenient to simulate breast cancer metastasis

Inactive Publication Date: 2019-11-22
HENAN UNIV OF CHINESE MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Most of the existing mouse models of breast cancer are primary sites, and there are few metastases at the primary sites. Breast cancer cells induced by a single method are not easy to metastasize to other parts. Therefore, it is not convenient to simulate the real mammary gland cancer metastasis

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] The modeling method of the bone metastasis mouse model of breast cancer of the present embodiment comprises the following steps:

[0021] 1) Making lentiviral preparations: the lentiviral vectors of lentiviral preparations are self-constructed or commercialized lentiviral vectors, which can be injected after packaging the lentiviral vectors, and the lentiviral expression plasmids and packaging plasmids pRev, pMDL, and pVSVG are co-transfected HEK293T cells were transfected, and the culture supernatant was collected 3 days after transfection, and the virus particles were obtained by centrifugal filtration. After diluting the virus, HEK293T cells were infected, and then the virus infection titer was tested.

[0022] 2) Make tumor cells: Stably transfect LacZ-labeled MCF-7 cells with the modified tet-transactivator protein in the bicistronic expression vector pBTE, and then use pTRE-hVEGF and a zeocin-resistant The plasmid pZeo-SV was used to retransfect the cells expressi...

Embodiment 2

[0028] The modeling method of the bone metastasis mouse model of breast cancer of the present embodiment comprises the following steps:

[0029] 1) Making lentiviral preparations: the lentiviral vectors of lentiviral preparations are self-constructed or commercialized lentiviral vectors, which can be injected after packaging the lentiviral vectors, and the lentiviral expression plasmids and packaging plasmids pRev, pMDL, and pVSVG are co-transfected HEK293T cells were transfected, and the culture supernatant was collected 3 days after transfection, and the virus particles were obtained by centrifugal filtration. After diluting the virus, HEK293T cells were infected, and then the virus infection titer was tested.

[0030] 2) Make tumor cells: Stably transfect LacZ-labeled MCF-7 cells with the modified tet-transactivator protein in the bicistronic expression vector pBTE, and then use pTRE-hVEGF and a zeocin-resistant The plasmid pZeo-SV was used to retransfect the cells expressi...

Embodiment 3

[0036] The modeling method of the bone metastasis mouse model of breast cancer of the present embodiment comprises the following steps:

[0037] 1) Making lentiviral preparations: the lentiviral vectors of lentiviral preparations are self-constructed or commercialized lentiviral vectors, which can be injected after packaging the lentiviral vectors, and the lentiviral expression plasmids and packaging plasmids pRev, pMDL, and pVSVG are co-transfected HEK293T cells were transfected, and the culture supernatant was collected 3 days after transfection, and the virus particles were obtained by centrifugal filtration. After diluting the virus, HEK293T cells were infected, and then the virus infection titer was tested.

[0038] 2) Make tumor cells: Stably transfect LacZ-labeled MCF-7 cells with the modified tet-transactivator protein in the bicistronic expression vector pBTE, and then use pTRE-hVEGF and a zeocin-resistant The plasmid pZeo-SV was used to retransfect the cells expressi...

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Abstract

The invention relates to a breast cancer bone metastasis mouse model molding method. The method comprises the following steps of sampling a mouse, performing nipple injection on the mouse through a slow virus preparation for inducing the mouse breast cancer, performing injection on the mouse obtained after nipple injection of the slow virus in a blood injection mode through the slow virus preparation for inducing the mouse breast cancer for several times. Firstly, the slow virus preparation for inducing the mouse breast cancer is used for performing nipple injection on the mouse, and the mousegenerates primary tumors. Then, the slow virus preparation for inducing the mouse breast cancer is used for performing blood injection on the mouse, primary tumor cells are induced to move in blood,and the bone metastasis state is formed. The primary breast cancer of the mouse is successfully transferred to the target locus.

Description

technical field [0001] The invention belongs to the technical field of mouse model modeling, and in particular relates to a method for making a mouse model of breast cancer bone metastasis. Background technique [0002] Breast cancer can spread to almost any area of ​​the body. The most common sites are lymph nodes, lung, bone, and liver. At present, breast cancer research is mainly based on breast cancer models established in rodents. Breast cancer mouse models are mainly used to simulate the occurrence, development and drug treatment of breast cancer, and provide relevant basis for clinical treatment of breast cancer. The existing animal models of breast cancer can be mainly divided into three categories: xenograft tumor models, physical and chemical carcinogen-induced tumor models, and transgenic models. Most of the existing mouse models of breast cancer are primary sites, and there are few metastases at the primary sites. Breast cancer cells induced by a single method...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/867A01K67/027
CPCA01K67/0271A01K2227/105A01K2267/0331C12N15/86C12N2740/15043
Inventor 韩倩倩王白燕高剑峰李宁杨联河姜乃菡
Owner HENAN UNIV OF CHINESE MEDICINE