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Inducing method and inducing liquid of mesenchymal stem cells

A technology of bone marrow mesenchyme and stem cells, applied in the field of cell engineering, can solve the problems of detachment, cell shrinkage, toxicity, etc., and achieve the effects of improving activity, improving transformation rate, and preventing detachment and apoptosis.

Inactive Publication Date: 2016-01-13
深圳华毓造血干细胞研究有限公司
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Problems solved by technology

However, after the cells are treated with 5-AZA, the cells can partially shrink, detach, degrade, or form fat vacuoles in the cytoplasm, indicating that 5-AZA, as an inducer, is also toxic to cells

Method used

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  • Inducing method and inducing liquid of mesenchymal stem cells
  • Inducing method and inducing liquid of mesenchymal stem cells

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Embodiment 1

[0045] The method for inducing bone marrow mesenchymal stem cells provided by the present invention comprises the following steps:

[0046] S1. Preliminary screening of bone marrow mesenchymal stem cells by density gradient centrifugation

[0047] Immediately inject the extracted bone marrow into a centrifuge tube containing 5ml of low-sugar DMEM and mix thoroughly. After filtering through a 200-mesh stainless steel filter, the mixed cell suspension is obtained, and the mixed cell suspension is gently added to the preset equal volume of Percoll In the centrifuge tube of the layered liquid (Percoll is a suspension of silica gel particles treated with polyvinylpyrrolidone, which is non-toxic and irritating to cells; derived from GE, the United States), the density of the Percoll layered liquid is 1.082kg / L. Centrifuge at 2400r / min for 20min, collect the mononuclear cells in cloudy buffy coat, wash twice with PBS solution at 1200r / min for 10min each time, and obtain preliminary s...

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Abstract

The invention relates to an inducing method of mesenchymal stem cells. The inducing method includes: obtaining the mesenchymal stem cells for in-vitro culture; using inducing liquid for induction in the process of in-vitro culture, wherein the inducing liquid comprises angiotensin II and bone morphogenetic protein-2. The bone morphogenetic protein capable of promoting growth and differentiation of the mesenchymal stem cells and the angiotensin II capable of promoting proliferation of the mesenchymal stem cells are adopted as inducers to induce culture of the mesenchymal stem cells, so that too-early wall-removal apoptosis of the mesenchymal stem cells can be prevented, and activity of the mesenchymal stem cells can be improved while conversion rate of the same can be increased.

Description

technical field [0001] The invention relates to the technical field of cell engineering, in particular to a method for inducing bone marrow mesenchymal stem cells and an inducing solution used. Background technique [0002] Coronary heart disease, myocardial infarction and heart failure are one of the major diseases that seriously affect people's quality of life and longevity. Research data show that mature cardiomyocytes lack regenerative ability. After myocardial infarction (MI), the myocardium in the infarcted area can only proliferate through fibrous tissue and be replaced by scar tissue without contractile function, resulting in a decline in cardiac function. Clinically, there is a lack of fundamental treatment methods for the regeneration and reconstruction of infarcted myocardium, and cell replacement therapy has emerged as an ideal treatment strategy. [0003] Bone marrow mesenchymal stem cells (Mesenchymaistemcells, MSCs) are a type of cells in the bone marrow simi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775
Inventor 曾宪卓鲁菲
Owner 深圳华毓造血干细胞研究有限公司
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