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A method for establishing a "human-derived" cardiac hypertrophy model based on pluripotent stem cell technology

A technology of pluripotent stem cells and cardiac hypertrophy, applied in the field of biomedicine, can solve the problems of lack of "human-derived" cardiac hypertrophy models

Active Publication Date: 2022-07-15
ZHEJIANG UNIV
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Problems solved by technology

[0004] The purpose of the present invention is to establish a method for establishing a "human-derived" cardiac hypertrophy model based on pluripotent stem cell technology, so as to solve the problem of lack of a "human-derived" cardiac hypertrophy model in the existing modeling technology, and to understand the mechanism of cardiac hypertrophy. Reliable human-derived models for research and development of therapeutic and preventive medicines

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  • A method for establishing a "human-derived" cardiac hypertrophy model based on pluripotent stem cell technology
  • A method for establishing a "human-derived" cardiac hypertrophy model based on pluripotent stem cell technology
  • A method for establishing a "human-derived" cardiac hypertrophy model based on pluripotent stem cell technology

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Embodiment Construction

[0021] 1) Directed differentiation of pluripotent stem cells to obtain cardiomyocytes: First, pluripotent stem cells such as H9 embryonic stem cells were seeded into a 6-well plate (Corning) at a ratio of 1:8-1:12, and the stem cell medium mTeSR (STEM CELL Technology) was replaced daily. ), when the cell density reaches about 80%, differentiation can be performed. On the 0th day, the old medium was aspirated, the myocardial differentiation medium (RPMI+B27-Insulin) (Gibco) was washed once, and 2 ml of differentiation medium containing 8 μM CHIR (Medchem Axon) was added to each well. After 2 days, it was replaced with myocardial differentiation medium. Differentiation medium, 2 ml per well, on the third day, 2 ml of myocardial differentiation medium containing 5 μM IWR (Medchem Axon) was added to each well, and the culture was continued for 2 days. Change to myocardial differentiation medium on day 5. After the 7th day, the medium was changed to cardiomyocyte medium (RPMI+B27+...

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Abstract

The invention discloses a method for establishing a "human-derived" myocardial hypertrophy model based on pluripotent stem cell technology. The method is to resuspend the cardiomyocytes differentiated from pluripotent stem cells that have been cultured for about 30 days and then inoculate them into a 12-well plate with 1.5 × 10 cells per well. 6 cell. About 24 hours after the cells adhere to the wall, the phorbol-12-myristate-13-acetate dosing experiment can be started, and the induction is carried out at a concentration of 1 μM for 4 days. The invention can establish a stable human-derived myocardial hypertrophy model based on the pluripotent stem cell technology, and the shape and function conform to the characteristics of myocardial cell hypertrophy, the operation is convenient, and the modeling success rate is high.

Description

Technical field: [0001] The invention belongs to the technical field of biomedicine, and relates to a method for establishing a "human-derived" myocardial hypertrophy model based on pluripotent stem cell technology, which can be used for mechanism research of myocardial hypertrophy and research and development of therapeutic and preventive drugs. Background technique: [0002] Hypertrophic cardiomyopathy is one of the most common heart diseases, with an incidence rate of 0.2-0.5% in the general population. There are at least two million patients with hypertrophic cardiomyopathy in my country, and the main pathological manifestation is myocardial hypertrophy. Cardiac hypertrophy is caused by various diseases that trigger increased myocardial workload, such as hypertension, valvular insufficiency, myocardial infarction, hereditary diseases, endocrine disorders, etc. Failure, malignant arrhythmia and even sudden cardiac death, seriously endanger human health. Models currently ...

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/077
CPCC12N5/0657C12N2509/00
Inventor 梁平唐玲王洪坤宫庭钰
Owner ZHEJIANG UNIV
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