Use of icariin in inducing external oriented differentiation of embryo stem cells

A technology of embryonic stem cells and icariin, which is applied in the field of pharmacology, can solve the problems of directional differentiation of embryonic stem cells that have not yet been seen with icariin

Inactive Publication Date: 2003-06-25
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0013] However, there is no report on the use of icariin to induce the directed differentiation of embryonic stem cells

Method used

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  • Use of icariin in inducing external oriented differentiation of embryo stem cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Example 1: Research on Icariin Induced Directed Differentiation of Embryonic Stem Cells (ES Cells) into Cardiomyocytes in Vitro

[0025] 1. Support culture of ES cells:

[0026] ES cell D3 cell line (ES-D3 cell) in the conditioned medium (containing DMEM medium, fetal bovine serum), press 2×10 7 -5×10 7 Cells / ml were inoculated on trophoblast cells (mouse fibroblasts or STO cell lines), and subcultured according to conventional methods.

[0027] 2. Icariin induced directed differentiation of ES cells into cardiomyocytes:

[0028] ① ES-D3 cells were digested with trypsin to make a single-cell suspension, inoculated on the inner surface of the lid of the culture dish in 20-30 μL, added 10ml D-Hanks solution into the culture dish, turned the lid over to form a hanging drop, and put it in the incubator Hanging drop culture for 2 days. Thereafter, the hanging drop was transferred to a petri dish filled with 10 ml of ES cell differentiation medium (containing DMEM medium ...

Embodiment 2

[0030] Example 2: The use of icariin in directed differentiation into cardiomyocytes in drug efficacy evaluation

[0031] If the embryoid somatic cell mass obtained in Example 1 can beat rhythmically, it will be used as an indicator of directed differentiation into cardiomyocytes, and the time-effect relationship curve and Dose-response curve (see figure 1 ), which is used as an indicator for drug efficacy evaluation. It can also be used for drug development and screening of active ingredients of traditional Chinese medicine. see figure 1 , the effect of icariin on inducing ES cells to differentiate into cardiomyocytes is obvious, and it has a good dose-effect relationship. When it is co-cultured with ES-D3 cells for 5-7 days, more than 90% of the cell clones appear rhythmic sexual beating, and the expression of cardiac germ fibrin was up-regulated. figure 1 Among them, the final concentrations of icariin were: -▲-0, -■-10 -8 , -◆-10 -7 mol L -1 . * P** P*** P-1 , each...

Embodiment 3

[0033] Example 3: Directed differentiation of icariin into cardiomyocytes in other applications

[0034] Using icariin to induce embryonic stem cells to differentiate into cardiomyocytes, if it can be successfully introduced into the failing heart and work with host cells, this pathway can become an important source of tissue cells for transplantation and treatment of cardiac insufficiency; The method can cultivate a large number of cardiomyocyte clones and provide tissue engineering and other research.

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Abstract

The present invention provides a new use icariin in inducing external oriented differentiation of embryo stem cells into single type of cells. Icariin may be used as the mitogen in stem cell transplanting cardiac muscle regeneration or cardiac muscle cell reconstruction, and in constructing high-efficiency pharmacodynamic screening model for initial screening and estimation of medicine effect. The present invention demonstrates the medicine effect of icariin in cell level and approaches its plentiful biological information. The present invention determines icariin as pharmacological active componnet and this provides Chinese medicine prevention and treatment with substantil foundation. At the same time, the description of the pharmacodynamic mechanism provides reference for modernized development of Chinese medicine.

Description

technical field [0001] The invention belongs to the field of pharmacology, and relates to the use of a traditional Chinese medicine monomer compound, in particular to the use of icariin in inducing embryonic stem cell directional differentiation in vitro, which can be used to promote cell regeneration in stem cell transplantation myocardial regeneration or cellular myocardial reconstruction. differentiating agent. Background technique [0002] Herba Epimedii (Herba Epimedii), also known as the fairy spleen, is the dry part of the ground of various plants of the Berberidaceae (Berberbidaceae) Epimedium (Epimedium). It is often used to treat impotence, dripping urine, weak waist and knees, coronary heart disease, chronic bronchitis, neurasthenia and poliomyelitis. Modern pharmacological research has proved that Epimedium contains special chemical components and significant biological activities, and is a drug with a wide range of pharmacological activities. [0003] Icariin ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61L27/00C07H17/065C12N5/0735
Inventor 楼宜嘉王志强王丹雁
Owner ZHEJIANG UNIV
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