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Process of producing cardiac cell from interval leaf stem cell

A stem cell, stem cell differentiation technology, applied in the field of producing cardiomyocytes

Inactive Publication Date: 2011-04-27
IND TECH RES INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0009] In summary, so far there is no published document describing how to use ascorbic acid to induce bone marrow mesenchymal stem cells to differentiate into cardiomyocytes, and there is an urgent need in the art for an improved method for producing cardiomyocytes from bone marrow mesenchymal stem cells. The method is not only fairly easy to use, but also produces differentiated cardiomyocytes in a short period of time to ensure its use in cell therapy

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  • Process of producing cardiac cell from interval leaf stem cell
  • Process of producing cardiac cell from interval leaf stem cell
  • Process of producing cardiac cell from interval leaf stem cell

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Embodiment 1

[0038] Isolation and culture of bone marrow mesenchymal stem cells (BMSCs)

[0039] 1.1 Isolation and culture of porcine bone marrow mesenchymal stem cells

[0040] Porcine bone marrow mononuclear cells were isolated and purified according to the method disclosed by Haynesworth et al. (Haynesworth et al., (1992) Bone 13(1), 81-8). Briefly, three Lee-sung mini-pigs (Luh et al., (2000) Transplantation 69(10), 2019-2027) (30-45 kg ) to draw bone marrow fluid from the posterior iliac spine. After diluting with HBSS (Gibco) at a ratio of 1:1, put it into a 50 ml centrifuge tube containing 1.073 g / ml Ficoll solution (Amersham), and centrifuge at 2,000 rpm for 40 minutes at room temperature. The mononuclear cells located between the upper layer and the solution interface were collected, and washed twice with HBSS (Gibco) solution again. Afterwards, the cells were cultured in α-MEM matrix (Hyclone) supplemented with 20% fetal bovine serum (Gibco, Lot No.: 1149239), 100 units / ml o...

Embodiment 2

[0045] Induces bone marrow mesenchymal stem cells to differentiate into cardiomyocytes

[0046] 2.1 Differentiation of BMSCs into cardiomyocytes induced by ascorbic acid

[0047] The BMSCs (1×10 4 cells / cm2) were cultured in α-MEM (Hyclone) supplemented with 20% fetal bovine serum (FBS) for 2 days, after which the medium was replaced with 1'MEM (Hyclone) supplemented with 10% FBS ( Gibco), and cultivated in this I'MEM for 1 day, after that, various differentiation agents were added, including 5-azacytidine (3uM), ascorbic acid (1mM), dimethyl sulfoxide (0.5%), or vitamin A(10 -9 M), to induce myotube formation. After 4 weeks of culturing with the above-mentioned differentiating agents, the appearance of differentiated mesenchymal stem cells can be confirmed by the formation of myotubes in the cytoplasm. Cultured in 5-azacytidine (Fig. 1A) and ascorbic acid (Fig. For mesenchymal stem cells under 1B), the cell differentiation ratios are about 10% (5-15%) and 25% (20-35%)...

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Abstract

The present invention reveals one method of generating cardiac muscle cell for medicinal use. One embodiment for generating cardiac muscle cell includes culturing mesenchymal stem cell in culture medium containing ascorbic acid. One other embodiment includes the first selecting fast growing mesenchymal stem cell population and the subsequent culturing the selected fast growing mesenchymal stem cell population in culture medium containing ascorbic acid.

Description

technical field [0001] The present invention relates to cardiomyocytes suitable for cell therapy and methods of producing said cells. More specifically, the present invention provides a method for producing cardiomyocytes comprising culturing bone marrow mesenchymal stem cells in a medium containing ascorbic acid. Background technique [0002] Mesenchymal stem cells (MSCs) are multipotent or limited-potential stem cells that can differentiate into at least one type of cells derived from the mesenchymal cell lineage, such as adipocytes, cardiomyocytes, and skeletal muscle cells (Galmiche et al. (1993) Blood 82:66-76; and Wakitani et al. (1995) Muscle Nerve 18:1417-1426). Therefore, more and more studies hope that MSCs can be used in the treatment of various diseases, abnormalities and disabilities, because MSCs can be obtained from autologous cells or tissues to provide sufficient renewal cell sources without immune rejection; and MSCs are not Like embryonic stem cells, MSC...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/06C12N5/08C12N5/071C12N5/074C12N5/0789
Inventor 陈婉昕林思佑
Owner IND TECH RES INST