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Method for directionally differentiating multipotential stem cell in vitro into myocardial cell

A technology for pluripotent stem cells and cardiomyocytes, applied in artificial cell constructs, animal cells, vertebrate cells, etc., can solve the problems of low safety, high price, limited production capacity, etc., and achieve high safety and simple and reliable methods. , the effect of low price

Active Publication Date: 2015-01-21
昆明圆梦生命科学研究院有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The adherent culture method adopts a planar 2D culture method, and stem cells are induced to differentiate in a monolayer growth mode, which greatly limits the production capacity and is not conducive to practical and industrial production
In addition, on the one hand, these methods require special substrates such as martrigel, laminin, etc.; on the other hand, various expensive protein growth factors such as ActivinA, BMP4, bFGF, etc. are required
These matrices and growth factors have obvious drawbacks: unknown ingredients, expensive, huge variance between different batches; some are of animal origin, resulting in low safety, etc.

Method used

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  • Method for directionally differentiating multipotential stem cell in vitro into myocardial cell
  • Method for directionally differentiating multipotential stem cell in vitro into myocardial cell
  • Method for directionally differentiating multipotential stem cell in vitro into myocardial cell

Examples

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Embodiment 1

[0031] Example 1 is used to establish a medium for inducing cardiomyocyte differentiation in vitro under three-dimensional suspension conditions

[0032] The medium provided in this example for establishing the differentiation of cardiomyocytes in vitro under three-dimensional suspension conditions includes: a differentiation medium for inducing the differentiation of pluripotent stem cells into mesoderm precursor cells; Differentiation medium for differentiation into cardiomyocytes and long-term maintenance medium for cardiomyocytes.

[0033] The differentiation medium used to induce differentiation of pluripotent stem cells into mesoderm precursor cells includes: the basal culture medium is a mixed medium of DMEM / F12 and IMDM, KOSR (serum substitute), β-mercaptoethanol, glutamine Amide, NEAA (non-essential amino acid), CHIR99021 (agonist of Wnt signaling pathway) and BIO (agonist of Wnt signaling pathway).

[0034] The differentiation medium that is used to differentiate me...

example 1

[0038] Example 1: DMEM / F12 was mixed with IMDM at a volume ratio of 1:2, 5% KOSR, 1% NEAA, 0.025% β-mercaptoethanol, 100 mM glutamine, 5 μM CHIR99021 and 5 μM BIO.

example 2

[0039] Example 2: DMEM / F12 was mixed with IMDM at a volume ratio of 1:1, 10% KOSR, 1% NEAA, 0.025% β-mercaptoethanol, 50 mM glutamine, 10 μM CHIR99021 and 10 μM BIO.

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Abstract

The invention provides a method for directionally differentiating a multipotential stem cell in vitro into a myocardial cell and provides a culture medium for establishing myocardial cell differentiation by means of induction in vitro under a three-dimensional suspension condition. The culture medium comprises a differential medium for induced differentiation of a multipotential stem cell into a precursor cell of mesoblast, a differential medium for differentiating the precursor cell of mesoblast into the myocardial cell and a long-term maintaining culture medium of the myocardial cell. The method provided by the invention is simple and reliable, low in cost, stable and efficient and high in safety. By adopting a suspension culture system, high-quality myocardial cells can be industrially produced without any subsequent screening and purifying steps, can be directly used for application demands of scientific research of heart development, cell therapy of heart diseases, transplanting due to cardiac trauma and drug screening, and has inestimable scientific, social and economical benefits.

Description

technical field [0001] The invention relates to the field of cell biology, in particular to a method for directional differentiation of pluripotent stem cells into cardiomyocytes in vitro under three-dimensional suspension conditions. Background technique [0002] Heart disease is a common disease that threatens human health, especially myocardial infarction caused by myocardial ischemia caused by coronary heart disease. There are 1.5 million cases in the United States and 2 million cases in China every year. After myocardial infarction, local myocardial cells in the ventricle will undergo necrosis, causing local scarring that cannot be repaired, causing cardiac arrhythmia and insufficient blood pumping, which will affect the patient's production and life in mild cases, and cause shock or death in severe cases. Threat to human health and social stability. But there is currently no good treatment. Stem cells have long-term self-renewal and the ability to differentiate into ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071
Inventor 李天晴蒋斌季维智
Owner 昆明圆梦生命科学研究院有限公司
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