In-vitro culture medium of myeloid mesenchyma stem cell and use thereof

A bone marrow mesenchymal and stem cell technology, applied in tissue culture, medical preparations containing active ingredients, microorganisms, etc., can solve problems such as limited cardiac function, limited ability of new blood vessels, and failure to reach them, so as to improve curative effect, Effect of improving viability and lumen formation ability

Active Publication Date: 2009-11-25
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the ability of MSCs to improve cardiac function is limited. A large amount of evidence shows that the improvement of cardiac function by MSCs transplantation is mild to moderate. Most research results show that the improvement of cardiac function by MSCs transplantation is only about 5-6%, and even some clinical trials The improvement of cardiac function was not observed, which is far from our expectations. The important reason is that most of the cells die within a short time after transplantation into the body. Studies have reported that 90% of MSCs are transplanted into the body. Death within 1 day after transplantation is also closely related to the limited neovascularization ability of MSCs after transplantation into harsh environments

Method used

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  • In-vitro culture medium of myeloid mesenchyma stem cell and use thereof
  • In-vitro culture medium of myeloid mesenchyma stem cell and use thereof
  • In-vitro culture medium of myeloid mesenchyma stem cell and use thereof

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Embodiment 1

[0019] 1. Under aseptic conditions, extract 30-50ml of bone marrow fluid from the posterior superior iliac spine of a suitable adult patient (excluding hematopoietic system diseases) (Li Moumou, 26 years old, from Hangzhou, I agree to donate bone marrow for research use), and place In a sterile glass bottle pre-added with 4000IU heparin, mix well to avoid coagulation of bone marrow fluid, and move to a laminar flow chamber biosafety cabinet for the next step.

[0020] 2. Add the same amount of low-sugar DMEM medium (adding 100U / ml penicillin and 100U / ml streptomycin) (purchased from GIBCO, USA) to dilute the bone marrow fluid into the sterile glass bottle where the bone marrow fluid is placed, and use a sterile pipette Fully blow off the bone marrow cells.

[0021] 3. Separation of bone marrow mononuclear cells according to the density gradient separation method. Add 20ml of lymphocyte separation medium (density 1.077±0.001g / ml, purchased from Shanghai Hengxin Chemical Reagen...

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Abstract

The invention provides a cell in-vitro culture medium for enhancing viability and angiogenesis ability after myeloid mesenchyma stem cell transplantation and a processing method thereof, wherein the culture medium is prepared by adding DMOG with the terminal concentration of 500 to 2000 [mu]mol / L to conventional cell culture medium; the method includes the followings: stable stem cell of myeloid mesenchyma after subculturing is disposed in the cell culture medium with the terminal concentration of DMOG of 500 to 2000 [mu]mol / L for cultivation for 24 to 36 hours, and the stem cell of myeloid mesenchyma after processing is used for cell transplantation. The invention has the advantage that the viability and angiogenesis ability of stem cell of myeloid mesenchyma can be efficiently improved by means of DMOG preprocessing so that the curative effect for rehabilitating tissue damage by stem cell transplantation of myeloid mesenchyma can be improved.

Description

(1) Technical field [0001] The invention relates to an in vitro medium for bone marrow mesenchymal stem cells with additives and a pretreatment method for enhancing the viability and angiogenesis ability of bone marrow mesenchymal stem cells (MSCs) after transplantation, so as to improve the ability of cell transplantation to repair tissue damage efficacy. (2) Background technology [0002] Nowadays, cardiovascular and cerebrovascular diseases have become the number one killer of human health. Although drug therapy and interventional therapy have developed rapidly in recent years, and the prognosis of patients with ischemic cardiomyopathy has been significantly improved, drug therapy and interventional therapy cannot repair necrosis after all. of the heart muscle. Bone marrow mesenchymal stem cells (MSCs) have the ability of self-renewal and lateral differentiation, and can differentiate into endothelial cells, vascular smooth muscle cells and cardiomyocyte-like cells after...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/08A61K35/28A61P9/10
Inventor 王建安刘先宝
Owner ZHEJIANG UNIV
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