Method for enhancing angiogenesis promoting capability of mesenchymal stem cells

A technology of mesenchymal stem cells and angiogenesis, applied in the field of stem cells and regenerative medicine, can solve the problems of limited ability of cytokines, low survival ability, and influence on treatment effect, so as to promote paracrine function, enhance blood flow reperfusion, and improve prophylaxis. The effect of angiogenesis ability

Pending Publication Date: 2022-07-12
麦迪森(江苏)医学研究有限公司
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  • Application Information

AI Technical Summary

Problems solved by technology

However, the ability of umbilical cord mesenchymal stem cells to express and secrete angiogenesis-related cytokines under normal culture conditions is limited, because the insufficient secretion of cytokines leads to poor therapeutic effect
Moreover, the local transplantation of umbilical cord mesenchymal stem cells under normal culture conditions into ischemic tissues of patients often has low survival ability due to the hypoxic environment of the tissues, which directly affects the later treatment effect.

Method used

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  • Method for enhancing angiogenesis promoting capability of mesenchymal stem cells
  • Method for enhancing angiogenesis promoting capability of mesenchymal stem cells
  • Method for enhancing angiogenesis promoting capability of mesenchymal stem cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1 Gene expression of UC-MSCs promoting angiogenesis-related cytokines under different conditions

[0028] 1. Select P4 generation UC-MSCs, inoculate them in 6-well plates at a density of 2*105 / cm2 after digestion, and culture them in serum-free medium (Beijing Youkang Hengye Biotechnology Co., Ltd. product number: NC0103+NC0103.S), randomly Divided into 3 groups, the normal group was cultured in an ordinary carbon dioxide incubator (5% CO2+20% O2), and the hypoxia group and the pretreatment group were cultured in a hypoxia incubator (5% CO2+5% O2);

[0029] 2. Change the medium after 24 hours, 2ml per well, normal group use serum-free medium for normal culture, and hypoxia group use pretreatment medium without ATRA (serum-free complete medium+VB2 5μM+VC 50μM+VA 0.05 μM + Coenzyme Q10 concentration of 1 μM) was cultured in a hypoxia incubator, pretreatment group used ATRA-containing pretreatment medium (serum-free complete medium + ATRA 5 μM + VB2 5 μM + VC 50 μM...

Embodiment 2

[0033] Example 2 Human Vascular Endothelial Cell Function Detection

[0034] 1. Select P4 generation UC-MSCs, inoculate them in 6-well plates at a density of 2*105 / cm2 after digestion, and culture them in serum-free medium (Beijing Youkang Hengye Biotechnology Co., Ltd. product number: NC0103+NC0103.S), randomly Divided into 3 groups, the normal group was cultured in a normal carbon dioxide incubator (5% CO2+20% O2), and the hypoxia group and the pretreatment group were cultured in a hypoxia incubator (5% CO2+5% O2);

[0035] 2. Change the medium after 24 hours, 2ml per well, normal group use serum-free medium for normal culture, and hypoxia group use pretreatment medium without ATRA (serum-free complete medium+VB2 5μM+VC 50μM+VA 0.05 μM + Coenzyme Q10 concentration of 1 μM) continued to culture in a hypoxic incubator, pretreatment group used ATRA-containing pretreatment medium (serum-free complete medium + ATRA 5 μM + VB2 5 μM + VC 50 μM + VA 0.05 μM + Coenzyme Q10 concentrat...

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Abstract

The invention provides a method for enhancing the angiogenesis promoting capability of mesenchymal stem cells, which is characterized in that the mesenchymal stem cells are cultured under a low-oxygen condition by adopting a pretreatment culture medium, and the pretreatment culture medium is prepared by adding stimulating factors into a serum-free culture medium, the stimulating factors comprise all-transretinoic acid, riboflavin, ascorbic acid, vitamin A and coenzyme Q10, and the oxygen content is 0.5%-10% under the hypoxia condition. According to the method, the umbilical cord mesenchymal stem cells are promoted to express and secrete angiogenesis-related cell factors including endothelial cell growth factors, angiogenin 1, basic fibroblast growth factors and the like by adopting a method of combining pretreatment culture medium with low-oxygen stimulation, so that the angiogenesis-promoting capability of the mesenchymal stem cells is improved, and the angiogenesis-promoting effect of the umbilical cord mesenchymal stem cells is improved. After transplantation, the promotion of host ischemic tissue capillary angiogenesis, the acceleration of collateral circulation reconstruction and the enhancement of blood flow reperfusion of the ischemic tissue are enhanced, and a better treatment method is provided for tissue regeneration and related diseases.

Description

technical field [0001] The invention belongs to the field of stem cells and regenerative medicine, and in particular relates to a method for enhancing the angiogenesis-promoting ability of mesenchymal stem cells. Background technique [0002] Mesenchymal stem cells (MSCs) are a group of adult stem cells from the mesoderm that have the ability to differentiate into various terminal cells such as chondrocytes, skeletal cells and adipocytes. In umbilical cord tissue, placental tissue, and adipose tissue. MSCs have a strong paracrine capacity. Studies have shown that normal cultured MSCs can secrete more than 200 cell growth factors to act on target cells and achieve characteristic physiological functions. Therefore, MSCs can regulate immune regulation, promote angiogenesis, and promote regeneration of damaged tissues. It supports important physiological functions such as hematopoiesis. It has been approved and marketed in many countries as a new drug for graft-versus-host dise...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775A61K35/28A61P9/10
CPCC12N5/0668A61K35/28A61P9/10C12N2500/90C12N2500/38C12N2500/30C12N2500/02
Inventor 朱建斌
Owner 麦迪森(江苏)医学研究有限公司
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