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Novel application of icariin

A technology for the application of icariin, which is applied in the field of new application of icariin, and can solve problems such as the limitation of differentiation ability

Inactive Publication Date: 2015-08-19
TIANJIN UNIV OF TRADITIONAL CHINESE MEDICINE +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Although hUCMSCs have certain stem cell characteristics, their differentiation ability still has certain limitations

Method used

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  • Novel application of icariin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0062] 1. Isolation, culture and identification of hUMSCs

[0063] Isolation of umbilical cord mesenchymal stem cells: Under sterile conditions, the umbilical cord was repeatedly washed with 2% double antibody D-hank's for 2-3 times to remove residual blood as much as possible and remove arterial and venous vessels. Place the cleaned umbilical cord in a glass dish and cut into 1mm 3 size, add collagenase I and collagenase II (final concentration: 0.05%), place in a constant temperature shaker at 37°C, and digest for 30 minutes. Filter through a coarse sieve, the filtrate is A, add 0.25% trypsin to the remaining tissue and continue to digest for 30 minutes; Mix liquid A and liquid B, then filter through a 200-mesh sieve, collect the filtrate, 1000rpm, 15min, add whole culture (DF12+20%FBS+1%L-G+1%PS+10ng / mlEGF+10ng / mlbFGF) , placed at 37°C saturated humidity, 5% CO 2 Incubator cultivation. The medium was changed twice a week, and the cells were subcultured when the cells re...

Embodiment 2

[0071] Effects of active ingredients of kidney-tonifying traditional Chinese medicine on the pluripotency of hUCMSCs

[0072] 1. Effects of active ingredients of different kidney-tonifying traditional Chinese medicines on the expression of pluripotency gene Oct-4 in hUCMSCs

[0073] The active ingredients of commonly used kidney-tonifying traditional Chinese medicines are selected: icariin, total flavonoids of Epimedium, total flavonoids of Cuscuta, polysaccharides from Morinda officinalis, naringin, loganin and morroniside, among which, polysaccharides from Morinda officinalis, total The flavonoids and the total flavonoids of Cuscuta were obtained by self-extraction, and the rest were purchased standard products. in,

[0074] The extraction method of Morinda officinalis polysaccharides is as follows: accurately weigh the medicinal material of Morinda officinalis, crush it, add 10 times the volume of water for ultrasonic extraction, recover the water extract and evaporate to ...

Embodiment 3

[0085] Effects of active ingredients of kidney-tonifying traditional Chinese medicine on the ability of umbilical cord mesenchymal stem cells to differentiate into three germ layer tissue cells

[0086] ⅠIn vitro study——The effect of hUMSCs on the differentiation ability of three germ layer tissue cells

[0087] 1. In vitro induction of hUCMSCs to differentiate into nerve cells: hUCMSCs of the 4th generation were used in the experiment, digested with 0.25% trypsin-0.02% EDTA, and the cells were sliced. After pre-induction with Lβ-mercaptoethanol in low-sugar DMEM (10% FBS) for 24 h, the pre-induction solution was sucked off, washed three times with PBS, and then washed with 2% dimethyl sulfoxide and a final concentration of 200 μmol / L butylated hydroxyanisole BHA. Serum-free and low-sugar DMEM / F12 was used for induction incubation, and the morphological changes of the cells were observed under an inverted microscope after 4-6 hours.

[0088] 2. Immunofluorescence cytochemical...

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Abstract

The invention provides a novel application of icariin. Expression of a pluripotent gene Oct-4mRNA can be improved after the icariin acts on an umbilical cord mesenchymal stem cell after a week, pluripotency of the umbilical cord mesenchymal stem cell is improved, the capacity of the umbilical cord mesenchymal stem cell differentiating a nerve cell, a myocardial cell and an islet cell can be improved, the methylation level of the pluripotent adjusting gene Oct-4 is reduced, and the acetylation level of histone H3K9 is promoted.

Description

technical field [0001] The invention relates to a new application of icariin. Background technique [0002] Stem cells have multi-directional differentiation potential and self-renewal ability, and have good application prospects in cell replacement therapy, gene therapy, tissue engineering, etc. In recent years, the basic and clinical research on stem cells has advanced by leaps and bounds, and people have a deeper understanding of stem cells. From the research of totipotent embryonic stem cells (ES cells), to the discovery of adult stem cells, to the induced pluripotent stem cells (iPS) obtained by people through somatic cell reprogramming, all reflect the wisdom and devotion of scientific researchers to stem cell research. hard work. People have expressed great enthusiasm and hope for the large-scale and routine clinical application of stem cells, but due to the constraints of immune rejection, tumorigenicity, ethical and moral issues and other factors of embryonic stem...

Claims

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Application Information

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IPC IPC(8): C12N5/0775
Inventor 边育红禇晓倩王丽齐新李文崔换天周会芳赵庆云武玉琳
Owner TIANJIN UNIV OF TRADITIONAL CHINESE MEDICINE
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