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A medium for inducing embryonic stem cells into cardiomyocytes and its application

A technology of embryonic stem cells and human embryonic stem cells, applied in animal cells, vertebrate cells, bone/connective tissue cells, etc., can solve the problem of unstable differentiation system

Active Publication Date: 2020-04-28
INST OF ZOOLOGY CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, rice recombinant human serum albumin is necessary, and the concentration is as high as 0.5mg / mL, and the difference between serum albumin batches will lead to the instability of the differentiation system

Method used

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  • A medium for inducing embryonic stem cells into cardiomyocytes and its application
  • A medium for inducing embryonic stem cells into cardiomyocytes and its application
  • A medium for inducing embryonic stem cells into cardiomyocytes and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0150] The preparation of embodiment 1 medium of the present invention

[0151] 1. prepare liquid culture medium of the present invention according to the component and proportioning that table 3 provides, and the step of described culture medium is as follows:

[0152] 1) Calculate the amount of RPMI-1640 dry powder required for the basic medium according to the total volume prepared;

[0153] 2) Dissolve RPMI-1640 powder directly in sterilized deionized water, stir evenly; add 2g of sodium bicarbonate per liter, and adjust the pH to 7.2 with concentrated hydrochloric acid. Then use 5M sodium chloride stock solution to increase the osmotic pressure of the basal medium to 295, filter through a 0.22 μm filter, and store at 4°C.

[0154] 3) According to the volume of the prepared 50× stock solution, according to 1mg / L L-carnitine, 0.5mg / L ethanolamine, 10mg / L putrescine, 0.01mg / L sodium selenite, 0.5mg / L The ratio of linoleic acid in L and transferrin in 3mg / L is weighed, di...

Embodiment 2

[0167] The preparation of embodiment 2 medium of the present invention

[0168] 1. prepare liquid culture medium of the present invention according to the component and proportioning that table 5 provides, and the step of described culture medium is as follows:

[0169] 1) Calculate the amount of RPMI-1640 dry powder required for the basic medium according to the total volume prepared;

[0170] 2) Dissolve RPMI-1640 powder directly in sterilized deionized water, stir evenly; add 2g of sodium bicarbonate per liter, and adjust the pH to 7.2 with concentrated hydrochloric acid. Then use 5M sodium chloride stock solution to increase the osmotic pressure of the basal medium to 300, filter through a 0.22 μm filter, and store at 4°C.

[0171] 3) According to the volume of the prepared 50× stock solution, according to the L-carnitine of 3mg / L, the ethanolamine of 2mg / L, the putrescine of 20mg / L, the sodium selenite of 0.02mg / L, the sodium selenite of 2mg / L The proportion of linole...

Embodiment 3

[0183] The preparation of embodiment 3 medium of the present invention

[0184] 1. prepare liquid culture medium of the present invention according to the component and proportioning that table 7 provides, and the step of described culture medium is as follows:

[0185] 1) Calculate the amount of RPMI-1640 dry powder required for the basic medium according to the total volume prepared;

[0186] 2) Dissolve RPMI-1640 powder directly in sterilized deionized water, stir evenly; add 2g of sodium bicarbonate per liter, and adjust the pH to 7.2 with concentrated hydrochloric acid. Then use 5M sodium chloride stock solution to increase the osmotic pressure of the basal medium to 300, filter through a 0.22 μm filter, and store at 4°C.

[0187] 3) According to the volume of the prepared 50× stock solution, according to the L-carnitine of 2mg / L, the ethanolamine of 1mg / L, the putrescine of 16mg / L, the sodium selenite of 0.016mg / L, the sodium selenite of 1mg / L The proportion of linol...

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Abstract

The invention provides a culture medium capable of inducing embryonic stem cells into myocardial cells. The culture medium is clear in chemical components, is a liquid culture medium with clear chemical components and comprises a basal culture medium RPMI-1640 and further comprises 1-3mg / L of L-carnitine, 0.5-2mg / L of ethanolamine, 10-20mg / L of putrescine, 0.01-0.02mg / L of selenite sodium, 0.5-2mg / L of linoleic acid, 3-6mg / L of transferrin, 50-200mg / L of vitamin C and / or vitamin C magnesium phosphate and 50-500upsilonM N-acetylcysteine. The invention also provides a preparation method and an application method of the culture medium. The culture medium is clear in chemical components, low in cost and very conductive to large-scale preparation and clinical application of myocardial cells.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a culture medium for inducing embryonic stem cells into cardiomyocytes. Specifically, the present invention relates to the use of a culture medium with fully defined chemical components to induce human embryonic stem cells to generate cardiomyocytes in a targeted and efficient manner; the present invention also relates to a method for inducing embryonic stem cells into cardiomyocytes and its application. Background technique [0002] Research surveys in recent years have shown that heart disease has always been at the top of the deadly diseases, and among heart diseases, myocardial infarction is the most important cause of death. In recent years, with the continuous improvement of people's material living standards, especially the intake of high-calorie and high-fat foods and lack of sufficient physical exercise, the incidence of myocardial infarction has been increasing year by year. ...

Claims

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Application Information

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IPC IPC(8): C12N5/077
Inventor 周琪韩鹏程谭元卿郝捷王磊王柳
Owner INST OF ZOOLOGY CHINESE ACAD OF SCI
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