Method for constructing human myocardial cell apoptosis model

A cardiomyocyte apoptosis and cardiomyocyte technology, which is applied in the field of constructing a human cardiomyocyte apoptosis model, can solve problems such as the lack of expression of hiPSC-CMAT2

Pending Publication Date: 2020-07-14
GUANGDONG BEATING ORIGIN REGENERATIVE MEDICINE CO LTD
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Problems solved by technology

Gene expression analysis of hiPSC-CMs revealed loss of AT2 expression in hiPSC-CMs, which may be responsible for the previously reported failure of angiotensin II to induce apoptosis in hiPSC-CMs

Method used

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  • Method for constructing human myocardial cell apoptosis model
  • Method for constructing human myocardial cell apoptosis model
  • Method for constructing human myocardial cell apoptosis model

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Embodiment Construction

[0037] The technical solution of the present invention will be further described below in combination with experiments.

[0038] 1. Obtain iPS cells according to conventional and recognized methods.

[0039] 2. Differentiate the iPS cells into cardiomyocytes and purify them according to conventional and recognized methods.

[0040] Or specifically, the following method can be used to obtain purified hiPSC-CM.

[0041] Acquisition of hiPSC-CM

[0042] 1.1 Culture of hiPSC cells: Human induced pluripotent stem cells DYR0100 (ATCC) were seeded on plates coated with Matrigel matrix (Corning, 354277), and then cultured with Stemflex medium (Gibco, A3349401). StemFlex Medium was changed every two days. iPSCs were passaged every 3 days or when the cell culture reached 80-90% confluency. During subculture, the cells were washed once with 1×DPBS (Gibco, 14040133), and then treated with 0.5 mM EDTA (Invitgen, 15575020) diluted with 1×DPBS (Gibco, 14190144) in a medium for 10 min a...

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Abstract

The invention discloses a method for constructing a human myocardial cell apoptosis model. The inventor accidentally finds that after angiotensin II with a certain concentration is used for treating hiPSC-CM for a certain time, the apoptosis of hiPSC-CM can be effectively induced. According to some examples of the invention, the angiotensin II is used for inducing the apoptosis of human myocardialcells, and the myocardial cell apoptosis model closer to the human body is constructed, so that a powerful tool and method are provided for subsequent pharmacological experiments.

Description

technical field [0001] The invention relates to a method for constructing a cell model, in particular to a method for constructing a human cardiomyocyte apoptosis model. Background technique [0002] Heart failure is a very prevalent chronic disease among the elderly, and its prevalence and incidence are increasing year by year. Nearly 26 million people worldwide are suffering from heart failure, with a 5-year survival rate of less than 53%. The annual global economic burden of chronic heart failure treatment exceeds $108 billion. Heart failure is the terminal manifestation of many cardiovascular diseases, eg, ischemic heart disease, rheumatic heart disease. Due to individual differences, it is difficult to achieve satisfactory results in the treatment of heart failure. It is urgent to establish methods to assess the effect of different drugs on each heart failure patient. [0003] With the development of iPS (induced pluripotent stem cell) technology and the establishme...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/077C12N5/10
CPCC12N5/0657C12N2510/00C12N2506/45C12N2501/32
Inventor 林彬高强庄建岑坚正麦锦连周丽诗
Owner GUANGDONG BEATING ORIGIN REGENERATIVE MEDICINE CO LTD
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