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Application of oxidized low density lipoprotein for inducing mesenchymal stem cell to differentiate to myocardium-like cell

A technology of low-density lipoprotein and bone marrow mesenchyme, which is applied in the direction of bone/connective tissue cells, animal cells, vertebrate cells, etc., and can solve the problems of high cost, large dosage, and low induction transformation efficiency

Active Publication Date: 2015-03-04
XINXIANG MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, when angiotensin II is used to induce the differentiation of BMMSCs into cardiomyocytes in vitro, there are problems such as large dosage, high cost, and low induction transformation efficiency, and angiotensin II is likely to cause side effects such as high blood pressure in vivo

Method used

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  • Application of oxidized low density lipoprotein for inducing mesenchymal stem cell to differentiate to myocardium-like cell
  • Application of oxidized low density lipoprotein for inducing mesenchymal stem cell to differentiate to myocardium-like cell
  • Application of oxidized low density lipoprotein for inducing mesenchymal stem cell to differentiate to myocardium-like cell

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Example 1: Isolation and cultivation of mouse bone marrow mesenchymal stem cells (BMMSCs)

[0020] Under sterile conditions, bone marrow mesenchymal stem cells were isolated from the femoral bone marrow of the mouse hindlimb, and added to DMEM medium containing 20% ​​fetal bovine serum at 5×10 5 / mL concentration inoculated in cell culture flasks, placed at 37 ° C, 5% CO 2 cultured in an incubator, and obtained high-purity BMMSCs after 3 weeks of culture (such as figure 1 shown).

Embodiment 2

[0021] Example 2: Induced Differentiation of Mouse Bone Marrow Mesenchymal Stem Cells (BMMSCs)

[0022] After 3 weeks of culture, BMMSCs were digested with 0.25% trypsin, washed with PBS buffer, then added to DMEM medium containing 20% ​​fetal bovine serum and mixed well; 6 BMMSCs were inoculated into culture flasks at a cell concentration of / mL, and 5 μg / mL of oxidized low-density lipoprotein (ox-LDL) was added to the medium, and then incubated at 37°C, 5% CO 2 The induction culture was carried out under the condition of , the medium was changed every 3 days and 5 μg / mL ox-LDL was added, and the differentiation was induced for 4 weeks.

Embodiment 3

[0024] Example 3: Uptake detection of dioctanoyltetramethylindocyanine perchlorate (Dil) fluorescently labeled ox-LDL (Dil-ox-LDL)

[0025] BMMSCs were divided into 1×10 5 / cm 2 The density was inoculated in a 12-well culture plate, and after 4 days of culture, 5 μg / mL of Dil-ox-LDL was added to the medium, and placed at 37 ° C, 5% CO 2 Incubate in an incubator for 30 minutes, then wash the cells twice with serum-free DMEM medium, observe and take pictures under a fluorescent microscope, the detection results are as follows figure 2 shown.

[0026] The detection results showed that BMMSCs had a strong ability to absorb ox-LDL.

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Abstract

The invention provides application of oxidized low density lipoprotein for inducing mesenchymal stem cells to differentiate to myocardium-like cells. The experiment shows that when oxidized low density lipoprotein with concentration of 5 micrograms / mL is added into a culture medium of mesenchymal stem cells, BMMSCs have remarkable cell proliferation, the expression of myocardium-like cell specific protein and myocardium cell indicative molecules can be detected, which means that the mesenchymal stem cells are directionally differentiated to myocardium-like cells, and the conversion rate of the myocardium-like cells is as high as 30%.

Description

technical field [0001] The invention relates to a method for inducing differentiation of bone marrow mesenchymal stem cells to cardiomyocytes, in particular to the application of oxidized low-density lipoprotein in inducing the differentiation of bone marrow mesenchymal stem cells to cardiomyocytes. Background technique [0002] Bone marrow mesenchymal stem cells (Bone Marrow Mesenchymal Stem Cells, BMMSCs) are a type of adult stem cells that exist in the bone marrow. They have strong self-renewal ability and multi-directional differentiation potential. , can differentiate into various adult cells such as osteoblast, cartilage, adipose, myoblast, nerve and endothelium. BMMSCs are currently one of the most widely used seed cells in tissue engineering and regenerative medicine, and have been clinically used in the treatment of nervous system and cardiovascular system diseases. Animal experiments have shown that BMMSCs can differentiate into functional cells of the tissues and...

Claims

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Application Information

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IPC IPC(8): C12N5/077
Inventor 林俊堂曹毓琳张芬熙王现伟李永海丰慧根连杰
Owner XINXIANG MEDICAL UNIV
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