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

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

Example Embodiment

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

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

[0061] Take adult mice, open the chest under aseptic conditions, cut 3 hearts, rinse the hearts with pH 7.4 PBS three times, then place them on ice, add 1 mL of DMEM with 0.08% pancreatin and 1% collagenase for digestion 50mL of DMEM digestion solution contains 2mL of 0.08% pancreatin and 2mL of 1% collagenase II (trypsin and collagenase II were ordered from Beijing Xinjingke Biotechnology Company and sigma Company) First use scissors and blades to cut the tissue Chop into 1-2mm pieces. Then add 5mL digestion solution (50mL DMEM digestion solution contains 2mL of 0.08% trypsin and 2mL of 1% collagenase II) for digestion, and digest in a shaker at a constant temperature for 4-6 minutes at...

Example Embodiment

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

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

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

[0069] After anesthesia, the mice were fixed on the experimental wooden board. ECG monitoring. Cut the trachea in the middle of the neck and intubate, connect the animal artificial ventilator, open the chest in the fourth intercostal space, carefully lift the pericardium and cut open, fully expose the heart and blood vessels. Between the pulmonary artery cone and the left atrial appendage, using the trunk of the left coronary vein as a 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 of the myocardium and exit next to the pulmonary artery Needle; After the ECG is s...

Example Embodiment

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

[0085] Six-week-old C57 mice were selected, and the myocardial infarction model mice were prepared according to the step (1) of Example 2. Five mice were used as the control group and five mice were used as the experimental group. When the model mice were prepared for 24 hours, the mice in the experimental group were injected with 1×10 at the myocardial infarction. 6 Mi-iPS cells. One week later and one month later, the area of ​​myocardial infarction in mice was changed and the degree of repair of myocardial function in mice was measured with a supersonic cardiograph. The change of mouse heart function (FS% value) after one month see image 3 .

[0086] image 3 The data provided in is an observation 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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