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Application of stem cell exosomes in promoting vascularization and angiogenesis

An angiogenesis and exosome technology, applied in animal cells, vertebrate cells, cardiovascular system diseases, etc., can solve problems such as bottlenecks in promotion, insignificant effects, and decreased proliferation and differentiation potential.

Inactive Publication Date: 2018-08-28
SHANGHAI CHANGHAI HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, stem cells have limitations such as lack of cell sources, secondary trauma, and time-consuming and laborious cell selection. In addition, mesenchymal stem cells are adult human cells, and their proliferation and differentiation potential decreases significantly after passage. Compared with the cost of extraction and preparation, the effect is not significant. Make its promotion into a bottleneck

Method used

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  • Application of stem cell exosomes in promoting vascularization and angiogenesis
  • Application of stem cell exosomes in promoting vascularization and angiogenesis
  • Application of stem cell exosomes in promoting vascularization and angiogenesis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] Example 1: Extraction and identification of exosomes derived from umbilical cord mesenchymal stem cells:

[0056] 1. Obtaining umbilical cord mesenchymal stem cells:

[0057] The umbilical cord came from the Obstetrics and Gynecology Department of Changhai Hospital. Primary cultured mesenchymal stem cells. After harvesting the umbilical cord, put it in a sterile Petri dish or bottle containing sterile PBS or DMEM medium, and operate as soon as possible. After repeated washing with phosphate buffer, soak in L-DMEM containing penicillin and streptomycin (concentration of penicillin and streptomycin is 100IU / ml) for about 5 minutes, and cut into tissue pieces with a diameter of about 1.5mm; containing 10% Fetal bovine serum L-DMEM nutrient solution, 5% CO 2 , Culture in saturated humidity at 37°C; after 7-10 days, the mesenchymal stem cells can be seen crawling out of the tissue, and the tissue pieces are removed and digested with 0.25% trypsin for subculture. Subseque...

Embodiment 2

[0064] Example 2: The effect of exosomes derived from umbilical cord mesenchyme on ischemic nonunion:

[0065] 1. Establish a mouse model of ischemic nonunion, model reference (J Orthop Res 2007; 25(1): 51-61). And set up uMSCs-Exo group (experimental group); uMSC group (positive control group); HEK293T-Exo group (negative control group); PBS group (blank control group).

[0066] 2. On the 1st and 7th days, the same dose (200 μg / ml) was injected into the nonunion part of the rat with a 1ml syringe. Samples were taken as needed on the 7th, 14th, 21st, and 31st days after injection.

[0067] 1) The vessel volume (Vessel Volume, VV) was detected by the three-dimensional reconstruction of μCT vessels, and the neovascularization and distribution around the callus were compared. The specific steps are as follows: After the rats were killed, a 20G catheter was retrogradely inserted into the left carotid artery and a small incision was made in the right jugular vein. Inject heparin...

Embodiment 3

[0069] Example 3: The effect of exosomes derived from umbilical cord mesenchymal stem cells on the vascularization of fat particles after free fat transplantation:

[0070] 1. Extraction and grouping of fat granules: Fat granules were collected from 10 cases of abdominal liposuction of female patients aged 22 to 35 in the Department of Plastic Surgery, Changhai Hospital Affiliated to Second Military Medical University. Coronary heart disease, hypertension, diabetes, infectious diseases, etc. were excluded. Talk to the patients before the operation and obtain their consent to obtain adipose tissue for experiments. After liposuction, the adipose tissue or particles were centrifuged at a low speed (200×g) for 5 minutes, and then the yellow fat suspension was removed. The fat volume calculation method is as follows: put the washed and centrifuged layer of yellow fat suspension (containing a large piece of fat tissue) into a 5ml syringe with a marked liquid level, trim the fat piec...

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Abstract

The invention relates to the technical field of biology, in particular to an application of exosomes from mesenchymal stem cell in preparation medicine of promoting vascularization or inducing angiogenesis. The exosomes, processed with gradient centrifugation, filtering and ultracentrifugation, is with high purity. The exosomes has the advantages of easy preparation and storage, high safety coefficient, easy absorption, wide medicine route, has targeting ability, and makes the endothelial cells of the blood vessels come together and create new vessels, so that the microcirculation is reconstructed to increase blood flow perfusion. The deficiency of other functions is not caused, and the exosomes has a wide application prospect.

Description

technical field [0001] The invention relates to the field of biotechnology, specifically, the application of stem cell exosomes in promoting vascularization and angiogenesis. It mainly involves the use of stem cell exosomes to promote vascularization of damaged parts or around grafts to repair or survive ischemic tissues or organs. Background technique [0002] In the case of many lesions, the body loses the ability to generate new blood vessels, resulting in tissue ischemia, fibrosis and even necrosis due to insufficient blood supply. For example, nonunion of broken ends after traumatic fracture; fat liquefaction and necrosis after autologous fat transplantation, ischemic gangrene of limbs caused by autologous peripheral artery disease, etc. Angiogenesis at the base is a crucial stage in the healing process. With the formation of new blood vessels, the blood supply at the base of the injury increases, which can provide sufficient oxygen, nutritional support, and growth fac...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K35/28A61P9/10C12N5/0775
Inventor 章云童方硕夏琰谢杨祝玮烨唐炜雅郝滋晨
Owner SHANGHAI CHANGHAI HOSPITAL
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