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Inhibitor for inducing pluripotent stem cells, and inducing method and application thereof

A multifunctional stem cell and inhibitor technology, applied in the field of pharmaceutical bioengineering, can solve the problems of increasing the risk of iPS clinical application, limiting iPS application, iPS cell carcinogenesis, etc., and achieving good structural stability, ability to pass through cell membrane, and induction efficiency. improved effect

Active Publication Date: 2013-01-23
INST OF ZOOLOGY CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0015] However, iPS cells induced by methods such as retroviruses and lentiviruses currently used have the risk of carcinogenesis
This greatly increases the risk of clinical application of iPS, and limits the application of iPS in the field of regenerative medicine, and may cause serious complications.

Method used

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  • Inhibitor for inducing pluripotent stem cells, and inducing method and application thereof
  • Inhibitor for inducing pluripotent stem cells, and inducing method and application thereof
  • Inhibitor for inducing pluripotent stem cells, and inducing method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0059] Example 1 microRNA-421 inhibitor induces iPS cells

[0060] In this example, cardiac fibroblasts were isolated from mouse heart tissue, and microRNA-421 inhibitors were used to induce the cardiac fibroblasts to generate iPS cells. The specific experimental steps are as follows:

[0061] Take an adult mouse, open the chest and cut three hearts under sterile conditions, rinse the heart three times with PBS with pH 7.4, then place it on ice, add 1 mL of DMEM with 0.08% trypsin and 1% collagenase to digest 50mL of DMEM digestion solution containing 2mL of 0.08% trypsin and 2mL of 1% collagenase II (trypsin and collagenase II were ordered from Beijing Xinjingke Biotechnology Co., Ltd. and sigma company respectively). Chop into small pieces of 1-2mm. Then add 5mL of digestion solution (50mL of DMEM digestion solution contains 2mL of 0.08% trypsin and 2mL of 1% collagenase II) for digestion, and at 37°C, digest at a constant temperature in a shaker for 4-6 minutes (the sh...

Embodiment 2

[0066] Example 2 Preparation and identification of mouse model of myocardial infarction

[0067] In this example, a mouse model of myocardial infarction was prepared and identified.

[0068] 1) The method of blocking the left coronary artery is used to induce myocardial infarction in mice, thereby establishing a mouse model of myocardial infarction.

[0069] After anesthesia, the mice were fixed on the experimental wooden board in the dorsal position. ECG monitoring. The trachea was incised in the middle of the neck and intubated, connected to the animal respirator, the chest was opened in the fourth intercostal space, the pericardium was carefully lifted and cut open to fully expose the heart and blood vessels. Between the conus pulmonary artery and the left atrial appendage, with the main trunk of the left coronary vein as the mark, insert the needle below the root of the left atrial appendage with a depth of 0.5mm; pass the 6 / 0 suture through the surface layer of the m...

Embodiment 3

[0084] Example 3 Using mi-iPS cells to treat myocardial infarction in mice, observe the changes in the size of myocardial infarction and the degree of cardiac function repair in mice.

[0085] Six-week-old C57 mice were selected to prepare myocardial infarction model mice according to step (1) of Example 2. With five mice as the control group and five mice as the experimental group, 1 × 10 6 mi-iPS cells. After 1 week and 1 month, the change of the area of ​​myocardial infarction in the mice and the recovery degree of the myocardial function of the mice were detected by using an ultrasonocardiometer. The change (FS% value) of mouse cardiac function after one month sees image 3 .

[0086] image 3 The data presented in are observations after one month.

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Abstract

The invention provides a method for preparing pluripotent stem cells by using a micro ribonucleic acid (RNA)-421 inhibitor, the prepared pluripotent stem cells, application of the inhibitor and the stem cells to preparation of medicines for treating heart diseases, and application of the inhibitor to in-vitro inducing of the stem cells. The carcinogenesis and tumorigenesis risk which is caused by inducing iPS cells by using virus can be avoided; and mi-iPS cells are induced by using the micro RNA inhibitor, and the micro RNA inhibitor is used for treating myocardial infarction of a mouse to ensure that the function of the mouse which suffers from myocardial infarction can be repaired, so that the carcinogenesis and tumorigenesis risk which is caused by inducing the iPS cells by using virus can be avoided. The myocardial infarction of the mouse is treated by using the micro RNA inhibitor for the first time, an obvious treatment effect is achieved, the micro RNA inhibitor has potential application value when developed into a medicine for treating the myocardial infarction of the mouse, and great innovation is provided.

Description

technical field [0001] The invention belongs to the field of medical bioengineering. In a nutshell, the present invention relates to a microRNA-421 inhibitor, the use of the inhibitor to induce pluripotent stem cells in vitro, a method for preparing pluripotent stem cells induced by the inhibitor, the stem cells produced by the method, the The use of the above-mentioned stem cells in the preparation of medicines or kits for treating heart diseases, and the use of the inhibitors in the preparation of medicines or kits for treating heart diseases, as well as the above-mentioned medicines for treating heart diseases Pharmaceutical preparations or kits for diseases. Background technique [0002] 1. Overview of Cardiovascular Diseases [0003] Cardiovascular disease is the number one killer of human health. About 17 million people die from cardiovascular disease worldwide every year, and more than half of them die from acute myocardial infarction. The mortality rate is close to...

Claims

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Application Information

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IPC IPC(8): C12N15/113C12N5/077A61K35/12A61K48/00A61P9/00A61P9/10A61P9/04A61K35/545
Inventor 李培峰李娜龙波王昆
Owner INST OF ZOOLOGY CHINESE ACAD OF SCI
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