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Method for differentiating human umbilical cord mesenchymal stem cells into myocardial-like cells

A technology of cardiomyocyte-like cells and mesenchymal stem cells, which is applied in the field of differentiation of human umbilical cord mesenchymal stem cells into cardiomyocyte-like cells, and can solve the problems of insufficient myocardial reconstruction and a small number of differentiated cells

Inactive Publication Date: 2015-12-23
王泰华
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0002] At present, stem cell therapy for cardiovascular diseases such as myocardial infarction and heart failure has been gradually carried out, but the adult stem cells used for heart disease transplantation are non-heart-derived, so the number of differentiation into cardiomyocytes and endothelial cells is very small, which cannot meet the needs of myocardial reconstruction. Therefore, finding a kind of pluripotent cardiovascular stem cells and increasing their differentiation rate has become an important part of stem cell transplantation in the treatment of cardiovascular diseases.

Method used

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  • Method for differentiating human umbilical cord mesenchymal stem cells into myocardial-like cells
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  • Method for differentiating human umbilical cord mesenchymal stem cells into myocardial-like cells

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Embodiment 1

[0040] The method for the differentiation of human umbilical cord mesenchymal stem cells to cardiomyocytes, the steps are as follows:

[0041] Step 1: Cell culture phase, as follows:

[0042] (1) Isolation and culture of umbilical cord MSCs: take a neonatal umbilical cord, put it into a culture plate, cut the umbilical cord along the inner side of the ligation, take the middle part, add physiological saline to wash the surface of the umbilical cord, and transfer it to another culture medium after the surface is clean and bloodless. In a dish, cut into small pieces of 2cm, peel off the umbilical cord epidermis, umbilical artery and umbilical vein to obtain Walton's jelly, wash it three times with PBS containing 1% double antibody, and cut it into 1mm with a scalpel 3 Tissue block, then inoculated on 75cm 2 Cultivate in a culture flask, the medium is α-MEM with a mass concentration of 10% FBS, observe the cell morphology with an inverted microscope, replace the medium after 72 ...

Embodiment 2

[0052] The method for the differentiation of human umbilical cord mesenchymal stem cells to cardiomyocytes, the steps are as follows:

[0053] Step 1: Cell culture phase, as follows:

[0054] (1) Isolation and culture of umbilical cord MSCs: take a neonatal umbilical cord, put it into a culture plate, cut the umbilical cord along the inner side of the ligation, take the middle part, add physiological saline to wash the surface of the umbilical cord, and transfer it to another culture medium after the surface is clean and bloodless. In a dish, cut into 2.5cm small sections, peel off the umbilical cord epidermis, umbilical artery and umbilical vein to obtain Walton's jelly, wash it three times with PBS containing 1% double antibody, and cut it into 1mm with a scalpel 3 Tissue block, then inoculated on 75cm 2 Cultivate in a culture flask, the medium is α-MEM with a mass concentration of 10% FBS, observe the cell morphology with an inverted microscope, replace the medium after 72...

Embodiment 3

[0064] The method for the differentiation of human umbilical cord mesenchymal stem cells to cardiomyocytes, the steps are as follows:

[0065] Step 1: Cell culture phase, as follows:

[0066] (1) Isolation and culture of umbilical cord MSCs: take a neonatal umbilical cord, put it into a culture plate, cut the umbilical cord along the inner side of the ligation, take the middle part, add physiological saline to wash the surface of the umbilical cord, and transfer it to another culture medium after the surface is clean and bloodless. In a dish, cut into small pieces of 3cm, peel off the umbilical cord epidermis, umbilical artery and umbilical vein to obtain Walton's jelly, wash it three times with PBS containing 1% double antibody, and cut it into 1mm with a scalpel 3 Tissue block, then inoculated on 75cm 2 Cultivate in a culture flask, the medium is α-MEM with a mass concentration of 10% FBS, observe the cell morphology with an inverted microscope, replace the medium after 72 ...

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Abstract

The invention relates to a method for differentiating human umbilical cord mesenchymal stem cells into myocardial-like cells. Compared with human embryonic stem cells and bone mesenchymal stem cells, the human umbilical cord mesenchymal stem cells from Whartonps jelly have obvious advantages that cost is low, the source is abundant, and no ethical issue is involved; compared with the bone mesenchymal stem cells, the multiplication capacity and differentiation capacity of the human umbilical cord mesenchymal stem cells do not decline along with increase of the passage number and the organismic age. In this way, the myocardial-like cells differentiated from the 5-Aza induced human umbilical cord mesenchymal stem cells are in an immature stage, still have multiplication capacity and can continue multiplication to form enough myocardial cells, which provides possibility for clinical application of the myocardial-like cells in cell transplantation treatment.

Description

technical field [0001] The invention belongs to the field of biological technology, and in particular relates to a method for differentiating human umbilical cord mesenchymal stem cells into cardiomyocytes. Background technique [0002] At present, stem cell therapy for cardiovascular diseases such as myocardial infarction and heart failure has been gradually carried out, but the adult stem cells used for heart disease transplantation are non-heart-derived, so the number of differentiation into cardiomyocytes and endothelial cells is very small, which cannot meet the needs of myocardial reconstruction. Therefore, finding a kind of pluripotent cardiovascular stem cells and improving their differentiation rate has become an important link in the treatment of cardiovascular diseases by stem cell transplantation. [0003] Some existing methods are listed as follows: [0004] 1) chemical reagent induction [0005] 5-azacytidine induced MSCs to differentiate into cells with card...

Claims

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Application Information

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IPC IPC(8): C12N5/077
Inventor 王泰华
Owner 王泰华
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