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Antiapoptosis high expression hVEGF165 (human Vascular Endothelial Growth Factor 165) cell model and building method thereof

A high-efficiency expression and cell model technology, applied in the field of biomedical research, can solve the problems of unsustainable high-efficiency expression and short culture life of BMSCs, and achieve the effect of prolonging the lifespan, improving the effect and wide application prospect.

Inactive Publication Date: 2012-01-25
THE SECOND AFFILIATED HOSPITAL OF GUANGZHOU MEDICAL UNIV
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Problems solved by technology

[0007] The first purpose of the present invention is to solve the short lifespan of BMSCs in vitro culture in the prior art and cannot continuously and efficiently express hVEGF 165 problem, providing a viable anti-apoptotic overexpression of hVEGF 165 cell model

Method used

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  • Antiapoptosis high expression hVEGF165 (human Vascular Endothelial Growth Factor 165) cell model and building method thereof
  • Antiapoptosis high expression hVEGF165 (human Vascular Endothelial Growth Factor 165) cell model and building method thereof
  • Antiapoptosis high expression hVEGF165 (human Vascular Endothelial Growth Factor 165) cell model and building method thereof

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specific Embodiment 1

[0039] Anti-apoptosis and high-efficiency expression of hVEGF involved in this example 165 The cell model, with purified rat bone marrow mesenchymal stem cells BMSCs as transfected cells, with hBCL-2 and hVEGF 165 Double-gene co-expression of recombinant adenovirus, hBCL-2 and hVEGF 165 Double-gene co-expression of recombinant adenovirus transfected purified rat bone marrow mesenchymal stem cells BMSCs, constituting anti-apoptosis and highly expressing hVEGF 165 cell model.

[0040] The establishment method and detection process of this embodiment include the following steps: 1) obtaining purified rat bone marrow mesenchymal stem cells BMSCs; 2) transfecting purified rat bone marrow mesenchymal stem cells BMSCs with recombinant adenovirus; 3) then, using The cell model established by rat bone marrow mesenchymal stem cells BMSCs with the highest transfection efficiency, anti-apoptosis and high expression of hVEGF 165 cell model.

[0041] The specific operation method is as ...

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Abstract

The invention relates to an antiapoptosis high expression hVEGF165 (human Vascular Endothelial Growth Factor 165) cell model and a building method thereof. The cell model takes BMSCs (Bone Mesenchymal Stem Cells) as transfection cells and coexpresses a recombinant adenovirus with double genes hBCL-2 and hVEGF165. The building method of the cell model comprises the following steps of: 1) separating and purifying rat BMSCs to obtain massive purified BMSCs with good vitality; 2) transfecting the BMSCs by the recombinant adenovirus, building the cell model built by the BMSCs transfected for 72 hours by adopting a multiplicity of infection (MOI) is equal to 400. The application characteristics are as follows: 1) the cell model is used for treating diseases such as myocardial infarction and the like; and 2) the cell model is used for being transplanted into a receptor organ. Compared with common BMSCs, the transfection recombinant adenovirus BMSCs provided by the invention has the advantages that cell apoptosis ratio is obviously reduced, service life is prolonged, the hVEGF165 can be efficiently expressed, the cell model can be conveniently transplanted to the receptor organ, and effect for treating diseases can be improved, thus the cell model and the building method thereof which are provided by the invention have wide application prospect.

Description

technical field [0001] The invention relates to an anti-apoptosis high-efficiency expression hVEGF 165 Cell models and methods for their establishment. It belongs to the technical field of biomedical research. Background technique [0002] Studies have shown that the angiogenic effect of VEGF is partly achieved by inducing the expression of BCL-2 in vascular endothelial cells. The high expression of BCL-2 induced by VEGF helps to improve the survival of vascular endothelial cells under conditions of hypoxia and poisoning [18] . Experiments by Xu et al. showed that BMSCs transplantation improved cardiac function after myocardial infarction mainly by parasecreting some cytokines such as VEGF and then up-regulating the expression of BCL-2 in cardiomyocytes to achieve cardioprotection. [19] . BCL-2 can also inhibit the apoptosis of vascular endothelial cells induced by NO and promote the formation of blood vessels [20] . Overexpression of BCL-2 in tumor cells promotes exp...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/10C12N5/0775C12N7/01
Inventor 陈敏生林育辉倪晓彬何晓青刘世明罗承锋
Owner THE SECOND AFFILIATED HOSPITAL OF GUANGZHOU MEDICAL UNIV
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