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Lentivirus-mediated cxcr7 high expression engineered endothelial progenitor cells for therapeutic application in ischemic diseases

A technology of endothelial progenitor cells and recombinant lentivirus, which is applied in the field of genetically modified endothelial progenitor cells, can solve the problems of inability to achieve sustained, stable and high expression of CXCR7 in EPCs, and achieve the effects of promoting capillary angiogenesis, enhancing capacity, and promoting circulation reconstruction

Active Publication Date: 2019-11-26
CHENGDU MEDICAL COLLEGE
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Problems solved by technology

[0003] Yan Xiaoqing, "CXCR7 Promotes Angiogenesis of Human Cord Blood-derived Endothelial Progenitor Cells", China Ph.D. Virus-transgenic endothelial cells (EPCs) have confirmed that high expression of CXCR7 can improve the survival ability and tube-like structure formation ability of EPCs, but because adenovirus can only achieve transient high expression of CXCR7, EPCs transplanted into the body need a longer period of survival , and can promote angiogenesis through paracrine or integration into the microvascular endothelium. Therefore, there are obvious problems in the transient high expression of CXCR7 for the treatment of ischemic diseases in EPC transplantation, and the sustained and stable high expression of CXCR7 in EPCs cannot be achieved.

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  • Lentivirus-mediated cxcr7 high expression engineered endothelial progenitor cells for therapeutic application in ischemic diseases
  • Lentivirus-mediated cxcr7 high expression engineered endothelial progenitor cells for therapeutic application in ischemic diseases
  • Lentivirus-mediated cxcr7 high expression engineered endothelial progenitor cells for therapeutic application in ischemic diseases

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Embodiment 1

[0028] Embodiment 1 Preparation of recombinant lentivirus and transgenic cells of the present invention

[0029] 1 Material method

[0030] 1.1 Main reagents

[0031] The pLVX-3FLAG-EGFP-Puro (pSB44) vector was purchased from Shanghai Shengbo Biotechnology Co., Ltd.; restriction endonucleases EcoR I, BamH I and T4 DNA ligase were purchased from China Dalian Bao Biological Company; plasmid pCMV-dR8.9 and pCMV-VSV-G were purchased from Addgene; plasmid extraction kit (Qiagen); 2×PCR MasterMix and DNA Marker were purchased from China Beijing Tiangen Biochemical Technology Co., Ltd.; DMEM high glucose (GIBCO); Opti-MEM ( GIBCO Company); EGM-2 was purchased from American Lonza Company; Fibronectin (FN) was purchased from Sigma Company; SunBio Trans-EZ (SunBio, Shanghai); primary antibody CXCR7 was purchased from abcam Company, Rabbit Anti-GAPDH was purchased from Shanghai Dr. German company, the secondary antibody GoatAnti-Rabbit IgG was purchased from Zhongshan Jinqiao Company; ...

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Abstract

The invention relates to application of a lentivirus-mediated CXCR7 high expression engineered endothelial progenitor cell in treatment of ischemic diseases, discloses a nucleotide sequence shown as SEQ ID NO.1, and a recombinant virus and a transgenic cell containing the sequence. The invention also discloses application of the transgenic cell. The invention constructs the new recombinant lentivirus expressing CXCR7 and the transgenic endothelial progenitor cell containing the recombinant lentivirus, the cell has significantly improved transendothelial migration ability, and tests prove that the cell can effectively promote angiogenesis, treat diabetic diabetes caused lower limb ischemia and cerebral ischemia-reperfusion injury, and has good clinical application prospects.

Description

technical field [0001] The invention relates to the field of cell transplantation therapy, in particular to a genetically modified endothelial progenitor cell. Background technique [0002] CXCR7, also known as RDC1 or CCX-CKR2, is a new receptor for stromal cell-derived factor-1 (SDF-1). CXCR7 is highly conserved in mammals. It is located on the second chromosome of human beings. It is located near the same chromosome as the CXCR1, CXCR2, and CXCR4 genes. It is widely expressed in tissues and organs such as the hematopoietic system, heart, bone, kidney, and brain. Existing studies have found that CXCR7 is closely related to angiogenesis. [0003] Yan Xiaoqing, "CXCR7 Promotes Angiogenesis of Human Cord Blood-derived Endothelial Progenitor Cells", China Ph.D. Virus-transgenic endothelial cells (EPCs) have confirmed that high expression of CXCR7 can improve the survival ability and tube-like structure formation ability of EPCs, but because adenovirus can only achieve transi...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/12C12N15/867C12N5/10A61K35/44A61P9/10
CPCA61K35/44C07K14/7158C12N5/0692C12N15/86C12N2510/00C12N2740/15043
Inventor 戴小珍晏小清谭毅张坤黄永
Owner CHENGDU MEDICAL COLLEGE
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