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Cardiac stem cell marker and application thereof

A technology of cardiac stem cells and markers, applied in the field of biomarkers, can solve problems such as absence of cardiac stem cells

Inactive Publication Date: 2017-12-08
THE FIRST AFFILIATED HOSPITAL OF SUN YAT SEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, there is no clear molecular marker on the surface of cardiac stem cells, so the identification of a biomarker of cardiac stem cells will provide new molecular markers for the isolation of endogenous cardiac stem cells, which is of great research significance

Method used

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  • Cardiac stem cell marker and application thereof
  • Cardiac stem cell marker and application thereof
  • Cardiac stem cell marker and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Embodiment 1: establish myocardial infarction model

[0019] All animal experiments in this example were approved by the Animal Ethics Committee.

[0020] experimental method:

[0021] 1) Coronary artery ligation: After anesthetizing the mouse, guide a flexible tube from the oral cavity into the trachea to connect to the ventilator, make an oblique incision on the left chest from the lower right to the upper left, and use a scalpel along the lower rib in the fourth intercostal space The intercostal muscles were incised to enter the thoracic cavity, and the heart was gently squeezed out. The proximal end of the left anterior descending artery was quickly sutured between the left atrial appendage and the conus pulmonary artery, and the proximal end of the left anterior descending branch was quickly sutured. At the same time, the changes of the electrocardiogram were observed. The syringe draws air out of the chest cavity to restore negative chest pressure. Mice with eje...

Embodiment 2

[0023] Embodiment 2: TNFR2-Bmx expression increases in human ischemic heart

[0024] The overall experimental technical route of the present invention is as attached figure 1 shown. The experimental method of this embodiment is to use anti-TNFR2 and anti-Bmx antibodies to perform IB or immunohistochemical detection of the expression of TNF receptor molecules and downstream effector molecules. The experimental samples are paraffin-embedded tissue sections obtained from Dr. Bradley of the University of Cambridge, UK during heart transplantation.

[0025] Its specific experimental method is:

[0026] 1) Immunofluorescence: tissue sections were fixed, Triton-X100 permeabilized, blocked with 1% BSA (bovine serum albumin) for 30 minutes, then rinsed with PBS (phosphate buffered saline), incubated with corresponding antibodies for 2 hours, rinsed with PBS, corresponding The fluorescent secondary antibody was incubated in the dark for 1 hour, the slide was mounted with glycerol aft...

Embodiment 3

[0030] Example 3: TNFR2 is co-expressed with Nkx2.5+ / Gata4+CSC markers in ischemic hearts

[0031] Wild-type, TNFR2-KO and Bmx-KO mice received a single subcutaneous injection of 5 mg kg-1 isoproterenol in the loose skin of the neck and were sacrificed at 1, 3, 6, 14 or 28 days, respectively (n=10 per group per time point). Cardiac function was detected by echocardiography before sacrifice. Use anti-TNFR2 and anti-Bmx antibodies for IB or immunohistochemical detection of TNFR2-Bmx expression. Bmx activation was detected by anti-phospho-Bmx. Mouse eCSCs were evaluated using anti-Gata4, anti-Nkx2.5. Simultaneously detect the expression of TNFR2 and Bmx and eCSC markers. Possible cardiac precursors were detected by immuno-double labeling for CSC markers (Gata4, NKx2.5 and α-SA).

[0032] The experimental results are attached image 3As shown in A, only some free cells are positive for Nkx2.5 and Gata4 in the normal heart. In the ischemic region of the ischemic heart, such ...

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Abstract

The invention provides a cardiac stem cell marker and application thereof. The cardiac stem cell marker contains TNFR2. According to the invention, the cardiac stem cell with positive TNFR2 is induced in ischemic myocardium of a mouse myocardial infarction model to be produced, a TNFR2-Bmx passage has the effect of activating ischemia-induced endogenous cardiac stem cells in the mouse myocardial infarction model, and the TNFR2 in an ischemic heart and a Nkx2.5+ / Gata4+CSC marker perform coexpression. In addition, the invention further provides a composition and a kit for screening the cardiac stem cell. The composition and the kit contain a reagent for detecting the TNFR2. Thereby, the TNFR2 serves as a new biomarker and provides a new molecular marker for separating the endogenous cardiac stem cells.

Description

technical field [0001] The present invention relates to a biomarker and its use, especially a biomarker related to cardiac stem cells and its use. Background technique [0002] Biomarkers refer to biochemical molecular indicators that can mark changes or possible changes in system, organ, tissue, cell and subcellular structure or function, and have a very wide range of uses. Biomarkers can be used for disease diagnosis, judgment of disease stage, or to evaluate the safety and effectiveness of new drugs or new treatments in the target population. [0003] For disease research, biomarkers generally refer to certain characteristic biochemical molecular indicators in a common physiological or pathological or therapeutic process that can be objectively measured and evaluated. Through its measurement, the current biological status of the body can be known. process in process. Examining a disease-specific biomarker may be helpful for disease identification, early diagnosis and pr...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/569
CPCG01N33/56966G01N33/6863G01N2333/525
Inventor 王敏纪卫东
Owner THE FIRST AFFILIATED HOSPITAL OF SUN YAT SEN UNIV
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