Method for preparing endothelial progenitor cell capturing bracket for gene modification and regulation

Abstract

The invention provides a method for preparing an endothelial progenitor cell capturing bracket for gene modification and regulation, which comprises the following steps: 1, preprocessing a bare bracket; 2, preparing an endothelial progenitor cell antibody bracket; 3, combining Caspase genes and specific promoter gene segments differentiated from smooth muscle cells in a gene recombination way andthen combining the composition of the Caspase genes and the specific promoter gene segments and eukaryotic expression vectors pEGFPC2 or pcDNA3.1 in the gene recombination way to obtain fusion genes;and 4, dissolving the fusion genes obtained in the step 3 in de-ionized water solution to obtain solution of which the concentration of the fusion genes is 10 to 100 mg / ml, placing the endothelial progenitor cell antibody bracket obtained in the step 2 into the solution for co-incubation at 4 DEG C for 12 minutes, taking the endothelial progenitor cell antibody bracket out, naturally airing the endothelial progenitor cell antibody bracket in a super clean bench for 2 to 6 hours and storing the endothelial progenitor cell antibody bracket at 4 DEG C. The method has the advantages of dual functions of resisting restenosis and protecting endothelial repair.

Description

technical field [0001] The invention provides a preparation method of a genetically modified and regulated endothelial progenitor cell capture scaffold, which belongs to the field of biotechnology. Background technique [0002] Coronary heart disease is one of the most common and serious diseases that endanger human health. Percutaneous coronary angioplasty (PTCA) and stent implantation have become the main means of interventional treatment of coronary heart disease. However, even drug-eluting stents cannot Fundamentally solve the problem of postoperative restenosis. The current drug-eluting stents inhibit the intimal proliferation of local blood vessels through the release of drugs, but at the same time inhibit the physiological healing process after vascular injury and slow down the repair of the endothelium, and the rapid healing of the endothelium is the inhibition of intimal smooth muscle cells. key to growth. Re-examining the mechanism of restenosis, it is precisely ...

AI Technical Summary

Problems solved by technology

Its efficacy has been found to have a certain effect on promoting endothelial healing in animal experiments and clinical trials, but at the same time, it has been reported that this type of scaffold can also non-specifically adsorb a large number of inflammatory cells while rapidly adsorbing EPCs. At the same time, EPCs in the body also have Some of them have the characteristics of smooth muscle precursor cells, so these cells may differentiate into smooth muscle cells after the scaffold is adsorbed, thereby aggravating restenosis. The risk of thrombosis cannot be completely avoided

At present, drugs such as rapamycin, paclitaxel and other cytotoxic drugs are usually used to inhibit restenosis, all of which have non-specific killing cells, thereby inhibiting the proliferation of endothelial cells and delaying endothelial repair.