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Preparation for promoting revascularization or angiogenesis and preparation method thereof

A kind of preparation, a new technology, applied in the direction of cardiovascular system diseases, medical preparations containing active ingredients, pharmaceutical formulas, etc., can solve the problems of the actual efficiency of surviving cells with low survival rate, and limit the promotion and application of stem cell therapy

Active Publication Date: 2011-01-12
盛泰英诺(嘉兴)医疗科技有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, a series of deficiencies in technology and application at this stage greatly limit the further promotion and application of stem cell therapy in clinical practice. These limitations include but are not limited to (1) the extremely low content of such stem cells in bone marrow and blood (approximately 0.01%); (2) the extremely large number of cells required for this type of cell therapy (1×105-1×106 cells / kg body weight); (3) the extremely low survival rate and surviving cells after cell transplantation into the body (less than 10%); and (4) dysfunction of the patient's own stem cells due to chronic disease and long-term effects of a large number of cardiovascular risk factors

Method used

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  • Preparation for promoting revascularization or angiogenesis and preparation method thereof
  • Preparation for promoting revascularization or angiogenesis and preparation method thereof
  • Preparation for promoting revascularization or angiogenesis and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1. Preparation of monocytes.

[0032] 100 mL of blood was added to the density gradient agent Histopaque-1077 (Sigma), and 30 mL of blood was added per 15 mL of Histopaque. Centrifuge the test tube containing the blood and the gradient reagent at 400G for 30 minutes at room temperature. After stratification, use a sterile disposable needle to draw the middle opaque layer, which is the suspension rich in mononuclear cells. Depending on the situation, every 100ml of blood can get 1x10 8 ~1x10 10 monocytes.

Embodiment 2

[0033] Example 2. Acquisition of EPC cells.

[0034] For the acquisition method of type I EPC cells: the mononuclear cell-rich suspension was added with 10% human serum albumin and 1% growth factor additive EGM (purchased by Swiss Lonza Company, which contains vascular endothelial growth factor VEGF-1, basic fibroblast growth factor FGF-2, epidermal growth factor EGF, and insulin-like growth factor IGF-1) in EBM-2 medium at 1×10 per square centimeter 6 The density of the cells was cultured for 4 days, and the adherent cells were digested with trypsin and collected at 2×10 per square centimeter. 5 density for 2 days. The obtained type I EPCs are spindle-shaped cells with typical endothelial cell characteristics of endocytosing acetylated-LDL and binding UEA-1 lectin, and expressing a large amount of vascular endothelial cells Cell growth factor receptors KDR, CD31, CD14, CD45, a small amount of expression of CD34, CD133 and VE-cadherin.

[0035] For the acquisition method of...

Embodiment 3

[0038] Example 3. Acquisition of pro-angiogenic or neoplastic agents.

[0039] The type I EPC cells obtained in Example 2 were placed in phosphate buffered saline (PBS) without any growth factors (2×10 per square centimeter). 5 cells), cultured in an environment with an oxygen concentration of 1.5% for 2 days, and supplemented with 1% medical human serum albumin. After the culture was completed, the cell-free medium rich in cell growth factors was collected, and the adherent EPC cells were discarded. The collected cell-free medium was passed through a filter with a pore size of 0.2 microns to remove cell impurities and debris, and then stored in a -80°C freezer for later use. As the case may be, every 1x10 6 Each EPC cell can prepare 2-5 ml of the angiogenic regeneration or neonatal preparation.

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Abstract

The invention relates to a preparation for promoting revascularization or angiogenesis and a preparation method thereof. The preparation method of the preparation comprises the following steps of: inducing mononuclear cells to obtain endothelial progenitor cells (EPCs); then culturing the EPCs under the conditions of low blood serum and low oxygen; and finally separating the EPCs to obtain an acellular culture medium, and carrying out relevant aftertreatment on the acellular culture medium to obtain the preparation. The experiments in vitro and vivo indicate that the preparation for promoting revascularization or angiogenesis can obviously promote the regeneration of vessel tissues or the restoration of damaged vessel tissues.

Description

technical field [0001] The invention relates to a preparation capable of promoting blood vessel regeneration or neogenesis and a preparation method thereof. Background technique [0002] Cardiovascular disease is the disease with the highest mortality and morbidity in the world. According to the statistical report of the American Heart Association in 2010, about 17.5 million people died of various cardiovascular diseases in the world in 2005, accounting for about 30% of the total number of deaths. It is estimated that by 2020, the number of deaths from cardiovascular diseases worldwide will reach 25 million. According to the WHO report, in the 10 years from 2006 to 2015, China's income loss due to cardiovascular disease and diabetes will be as high as 558 billion US dollars. In 2004, 4.3 million people died of various cardiovascular diseases in Europe, accounting for 48% of the total number of deaths. Among various cardiovascular diseases, thrombus and vascular blockage c...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K35/14A61K35/28C12N5/0789A61P9/00A61P9/10
Inventor 杨肖泱杨子江顾丽娅
Owner 盛泰英诺(嘉兴)医疗科技有限公司
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