Preparation method of tumor suppressor gene-modified mesenchymal stem cells

Abstract

The invention discloses a preparation method for tumor suppressor gene-modified mesenchymal stem cells. The method comprises the following steps: A) separation of MSCs and primary culture and subculture; B) building of a tumor suppressor gene recombinant vector; and C) expression and identification of the tumor suppressor gene in MSCs. According to the method, recombination of a target tumor suppressor gene is carried out by adopting an adenovirus vector, the target gene is jointly expressed in the MSCs after the recombinant plasmid transfects the MSCs, and the preparation method has high infection rate and coexpression efficiency. Compared with other virus vectors, the vector disclosed by the invention has the advantages that the adenovirus transduction efficiency is relatively high, and fissional stem cells can be efficiently transfected, and the vector is not integrated into chromosome. Therefore, the safety is relatively high.

Description

[0001] technical field [0002] The invention belongs to the technical field of preparation of mesenchymal stem cells, in particular to a method for preparing mesenchymal stem cells modified by tumor suppressor genes. [0003] Background technique [0004] Gene therapy refers to a new biomedical technology that introduces genes with normal functions or genes with therapeutic effects into target cells in a certain way to correct gene defects or exert therapeutic effects, so as to achieve the purpose of treating diseases. The principle of tumor gene therapy is to introduce the target gene into target cells with gene transfer technology, so that it can obtain specific functions, and then execute or mediate the killing and inhibition of tumors, or protect normal cells from the serious effects of chemotherapy and radiation therapy. harm. According to different treatment strategies, the target gene can be proto-oncogene or antisense nucleic acid of tumor autocrine growth factor ...

AI Technical Summary

Problems solved by technology

Viruses have safety risks, such as causing insertion mutations, forming anti-DNA antibodies, local infection, and causing tumor nodules and ulcers, etc., and their immunogenicity also limits repeated administration

Non-viral gene targeting delivery carriers, such as liposomes, magnetic nanoparticles, cationic polymers, and polymer vesicles, have low immunogenicity and good biocompatibility, but there are still shortcomings, such as The gene transfer efficiency mediated by non-viral vectors is low, the particle size distribution of magnetic nanoparticles is uneven, and it is easy to form aggregates to block capillaries. Liposomes themselves are rapidly recognized and cleared in the bloodstream by the reticuloendothelial system, limiting their further application as gene carriers

Therefore, the safety, targeting, low transfection efficiency, and short expression time of existing viral and non-viral gene carriers limit the development and application of tumor gene therapy. Currently, it is necessary to develop safer and more efficient gene carriers to support Related scientific research and clinical application of tumor gene therapy

Images