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MiR-17-92 modified mesenchymal stem cell, miR-17-92 modified mesenchymal stem cellexosome, preparation methods for miR-17-92 modified mesenchymal stem cell and miR-17-92 modified mesenchymal stem cellexosome and application of miR-17-92 modified mesenchymal stem cell and miR-17-92 modified mesenchymal stem cellexosome

A technology of mir-19a and mir-19b, applied in the field of mesenchymal stem cell genetic modification, can solve problems such as changing the composition and abundance of exosomes, and achieve the effects of improving migration ability, promoting skin wound healing, and improving survival rate.

Pending Publication Date: 2021-01-22
JILIN UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Exosomes derived from stem cells only have the biological characteristics of stem cells. Therefore, if the composition and abundance of exosomes can be changed, the effect of exosomes can be enhanced, so that exosomes from stem cells can better promote angiogenesis, Regulate immunity, inhibit fibrosis, promote cell proliferation, migration and differentiation; currently there is no effective method to change the composition and abundance of exosomes

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  • MiR-17-92 modified mesenchymal stem cell, miR-17-92 modified mesenchymal stem cellexosome, preparation methods for miR-17-92 modified mesenchymal stem cell and miR-17-92 modified mesenchymal stem cellexosome and application of miR-17-92 modified mesenchymal stem cell and miR-17-92 modified mesenchymal stem cellexosome
  • MiR-17-92 modified mesenchymal stem cell, miR-17-92 modified mesenchymal stem cellexosome, preparation methods for miR-17-92 modified mesenchymal stem cell and miR-17-92 modified mesenchymal stem cellexosome and application of miR-17-92 modified mesenchymal stem cell and miR-17-92 modified mesenchymal stem cellexosome
  • MiR-17-92 modified mesenchymal stem cell, miR-17-92 modified mesenchymal stem cellexosome, preparation methods for miR-17-92 modified mesenchymal stem cell and miR-17-92 modified mesenchymal stem cellexosome and application of miR-17-92 modified mesenchymal stem cell and miR-17-92 modified mesenchymal stem cellexosome

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preparation example Construction

[0034] The invention provides the preparation method of the miRNA-17-92 modified mesenchymal stem cells, comprising the following steps: 1) isolating and culturing the mesenchymal stem cells; 2) using a lentivirus carrying the miR-17-92 gene cluster Transfect the mesenchymal stem cells described in step 1) to obtain miRNA-17-92 modified mesenchymal stem cells.

[0035] In the present invention, the mesenchymal stem cells are preferably human umbilical cord mesenchymal stem cells. In the present invention, the human umbilical cord mesenchymal stem cells are derived from fresh umbilical cords of normal healthy fetuses; the fresh umbilical cords are preferably obtained under sterile conditions; informed consent. In the present invention, after obtaining the fresh umbilical cord, preferably under aseptic conditions, the adventitia of the umbilical cord is peeled off, the umbilical artery and umbilical vein are removed, and after being fully rinsed with PBS, the umbilical cord mes...

Embodiment 1

[0046] (1) Isolation and culture of primary human umbilical cord mesenchymal stem cells

[0047] Fresh umbilical cords of normal healthy fetuses were obtained under aseptic conditions with the permission of the Ethics Committee and the informed consent of the puerpera. The adventitia of the umbilical cord was stripped in an ultra-clean workbench, the umbilical artery and umbilical vein were removed, and after being fully washed with PBS, the umbilical cord mesenchyme was cut into pieces and transferred to a sterile centrifuge tube, and the primary cultured human umbilical cord mesenchyme was digested and cultured with type I collagenase. Mesenchymal Stem Cells (UC-MSCs).

[0048] The concentration of type I collagenase is 1.0mg / ml; The volume ratio of type I collagenase and umbilical cord mesenchyme is 1:1; Put into cell culture box (5% CO 2 , 37°C) for 3-7 hours to obtain the cell suspension. After being inoculated into a 10cm cell culture dish and cultured for 3 generation...

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Abstract

The invention provides an miR-17-92 modified mesenchymal stem cell, an miR-17-92 modified mesenchymal stem cellexosome, preparation methods for the miR-17-92 modified mesenchymal stem cell and the miR-17-92 modified mesenchymal stem cellexosome and application of the miR-17-92 modified mesenchymal stem cell and the miR-17-92 modified mesenchymal stem cellexosome, and belongs to the technical fieldof mesenchymal stem cell gene modification. The mesenchymal stem cell is obtained through transfection with a lentivirus carrying a miR-17-92 gene cluster; The preparation method for the miRNA-17-92modified mesenchymal stem cell comprises the following steps: 1) separating and culturing the mesenchymal stem cell; and 2) transfecting the mesenchymal stem cell in the step 1) with the lentivirus carrying the miR-17-92 gene cluster to obtain the miRNA-17-92 modified mesenchymal stem cell. The miRNA-17-92 modified mesenchymal stem cell exosome disclosed by the invention can remarkably improve thesurvival rate of human umbilical cord mesenchymal stem cells, and improve the migration capability of the human umbilical cord mesenchymal stem cells; and the the miR-17-92 modified mesenchymal stemcell and the miR-17-92 modified mesenchymal stem cellexosome can be applied to preparation of drugs for promoting skin wound healing.

Description

technical field [0001] The invention belongs to the technical field of genetic modification of mesenchymal stem cells, and in particular relates to a mesenchymal stem cell modified by miRNA-17-92, exosomes and a preparation method and application thereof. Background technique [0002] The skin is often damaged by various internal pathologies and external mechanical factors. These acute and chronic skin defects have brought heavy burdens to patients and society. How to accelerate wound healing and promote wound repair is particularly important. Skin damage is the comprehensive result of the interaction of multiple cell death modes and inflammatory factors, and the treatment of skin damage should be a comprehensive regenerative strategy. [0003] In the field of wound repair, mesenchymal stem cell therapy has achieved gratifying therapeutic effects. Initial studies believed that the role of mesenchymal stem cells in wound repair was mainly related to their multi-lineage diff...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/10C12N15/113C12N15/867A61K35/28A61P17/02A61P43/00
CPCC12N5/0665C12N15/113C12N15/86A61K35/28A61P17/02A61P43/00C12N2510/00C12N2310/141C12N2740/15043
Inventor 王立生聂文博孙慧燕赵丽晶张丹
Owner JILIN UNIV