Method of testing amentoflavone in preparation of bone tissue engineering seed cells

A kind of technology of biflavone and bone tissue engineering, which is applied in the experimental field of biflavone of spiconia in the preparation of seed cells for bone tissue engineering, to achieve the effect of promoting early osteogenic differentiation, promoting MSCs proliferation, and promoting late osteogenic differentiation

Active Publication Date: 2015-12-23
GUANGDONG MEDICAL UNIV
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  • Summary
  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

However, there are no reports at home and abroad whether the biflavonoids can promote the osteogenic differentiation of MSCs.

Method used

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  • Method of testing amentoflavone in preparation of bone tissue engineering seed cells
  • Method of testing amentoflavone in preparation of bone tissue engineering seed cells
  • Method of testing amentoflavone in preparation of bone tissue engineering seed cells

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Embodiment Construction

[0025] The following will clearly and completely describe the technical solutions in the embodiments of the present invention with reference to the accompanying drawings in the embodiments of the present invention. Obviously, the described embodiments are only some, not all, embodiments of the present invention. Based on the embodiments of the present invention, all other embodiments obtained by persons of ordinary skill in the art without making creative efforts belong to the protection scope of the present invention.

[0026] A test method for preparing seed cells of bone tissue engineering with biflavonoids.

[0027] (1) MSCs culture

[0028] MSCs were placed in α-MEM medium containing 10% fetal bovine serum at 37°C, 5% CO 2 Cultivate in the incubator, change the medium once every 2-3 days, when the cells reach 80% confluence, digest and subculture with 0.25% trypsin;

[0029] (2) Cell proliferation detection

[0030] MSCs cells were seeded in 96-well culture plates, and...

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Abstract

The invention discloses a method of testing amentoflavone (AME) in the preparation of bone tissue engineering seed cells. The method includes: culturing mesenchymal stem cells (MSCs), adding osteogenesis-inducing medium (OIM) or osteogenesis-inducing medium containing different concentrations of amentoflavone, and detecting cell proliferation by an MTT (methyl thiazolyl tetrazolium) method; after the cells completely fuse, adding osteogenesis-inducing medium or osteogenesis-inducing medium containing different concentrations of amentoflavone to carry out culturing, adding a cell lysis solution, detecting activity of alkaline phosphatase with a kit, and measuring protein concentration by a Bio-Rad method; staining the cells with the alkaline phosphatase, and observing and photographing the cells under a microscope; after the cells completely fuse, adding osteogenesis-inducing medium or osteogenesis-inducing medium containing different concentrations of amentoflavone into each hole, and measuring osteocalcin level with an osteocalcin detection kit.

Description

technical field [0001] The invention relates to the field of bone tissue cell culture, in particular to a test method for the preparation of seed cells of bone tissue engineering by abiflavonoids of spicanthus spicosa. Background technique [0002] Bone defects caused by infection, tumor or trauma are common clinical diseases. The commonly used clinical methods for repairing bone defects include bone grafting and artificial bone, etc., but they all have certain defects and cannot meet the clinical needs. Bone tissue engineering provides a good prospect for bone defect repair. Bone tissue engineering must have suitable seed cells and scaffold materials. Seed cells for bone tissue engineering should meet the following requirements: easy to obtain materials, sufficient sources, and little damage to the body; have osteogenic potential; easy to expand in vitro and good biocompatibility. MSCs have been widely used as seed cells for bone tissue engineering, but how to induce ost...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775C12N5/077
Inventor 徐道华周晨慧张雪
Owner GUANGDONG MEDICAL UNIV
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