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Preparation method of stem cell exosomes and novel application thereof

An exosome and stem cell technology, applied in the field of preparation of stem cell exosomes, can solve the problems affecting the living standard and quality of patients, lack of compensatory collaterals, lower extremity arterial occlusion, etc., to provide survival rate and improve microcirculation. , the effect of promoting angiogenesis

Inactive Publication Date: 2019-09-03
广州市番禺区中心医院
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

At present, there is still a lack of effective treatment methods for diabetic lower extremity vascular disease. Clinically, drugs, interventional surgery and other methods are generally used for treatment, but these treatment methods have certain limitations, especially the occlusion of the lower extremity arterial outflow tract and the lack of compensatory collaterals. Formation, it is basically impossible to find suitable treatment to recanalize blood vessels [1] , the patient can only choose amputation, which will not only seriously affect the patient's living standard and quality, but also bring serious psychological trauma

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  • Preparation method of stem cell exosomes and novel application thereof
  • Preparation method of stem cell exosomes and novel application thereof

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Embodiment 1

[0029] A method for preparing exosomes from mesenchymal stem cells: take out mesenchymal stem cells (MSCs) from a low-temperature container, quickly place them in a 37°C water bath for continuous shaking, and quickly thaw within 1 min; under sterile conditions, completely thaw Transfer the cell suspension to a centrifuge tube containing 10ml of serum-free cell culture medium; centrifuge at 1500rpm for 5min, and discard the supernatant after centrifugation; use the medium to adjust the cell seeding density to 1.0×10 4 live cells / cm2, add the cell suspension to the culture vessel, shake gently to distribute the cells evenly; observe the state of the cells, and change the medium every other day.

[0030] Stem cell exosome isolation was performed at 4 °C: 2000 g for 10 min, 10,000 g for 20 min. Finally, stem cell exosomes were isolated by ultracentrifugation at 100,000 g for 120 min in a Beckman Coulter ultracentrifuge. The pelleted stem cell exosomes were collected and resuspend...

Embodiment 2

[0032] The new application of exosomes in the treatment of diabetic foot with stem cell exosomes combined with arterial endovascular angioplasty

[0033] Concrete embodiment:

[0034] (1) After systemic heparinization was given through the vascular sheath, a 0.035in (1in=0.0254m) spade guide wire was inserted, and the catheter reached the proximal end of the occlusion site.

[0035] (2) Routine arteriography of the affected extremity throughout the whole process is required to show the best microcirculation of the foot (the total amount of contrast agent is 18ml, and the speed is 4ml / mim). According to the intraoperative development, the length of stenosis and the degree of lesion were determined (the total amount of contrast medium was 14ml, and the speed was 3ml / mim). The speed and dose can be better observed.

[0036] (3) Arterial endovascular angioplasty is performed at the arterial occlusion, and a catheter is placed, and the patient can return to the ward. Subsequentl...

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Abstract

The invention relates to the field of biological preparations, in particular to a preparation method of stem cell exosomes and a novel application thereof. The method comprises the steps of extractionof mesenchymal stem cell exosomes, percutaneous transluminal coronary angioplasty, and stem cell exosome transplantation, and can be used for treating diabetic foot. The novel application of exosomescan accurately and effectively improve the microcirculation of diabetic foot, promotes angiogenesis, reconstructs capillary network, reduces occlusion of blood vessel restenosis, reduces amputation rate, and provides survival rate.

Description

technical field [0001] The invention relates to the field of biological preparations, in particular to a preparation method and new application of stem cell exosomes. Background technique [0002] In recent years, with the aggravation of population aging and changes in people's diet structure, the incidence of diabetes has risen rapidly. Diabetes-induced ischemic diseases of the lower limbs (commonly known as "diabetic foot") can lead to amputation and even life-threatening. It has been reported that 15% of diabetic patients will have diabetic foot, the amputation rate of diabetic foot is about 25-50%, the amputation rate is 40 times that of normal people, and the 5-year survival rate is about 50%. Therefore, diabetic foot has attracted more and more attention. [0003] Diabetic lower extremity vasculopathy is a complex pathophysiological process, and its pathogenesis has not been fully elucidated. Most studies support the hypothesis that diabetic foot is a chronic low-grad...

Claims

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Application Information

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IPC IPC(8): A61K35/28A61K9/08A61P3/10A61P17/02A61P9/10A61P7/02A61P9/14C12N5/0775
CPCA61K35/28A61K9/08A61K9/0019A61P3/10A61P17/02A61P9/10A61P7/02A61P9/14C12N5/0662C12N2509/00
Inventor 黄晨罗文峰唐郁宽陈汉威庄炜钊李伟奇贺雪平
Owner 广州市番禺区中心医院
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