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Osteogenic induction medium and osteogenic differentiation method

A technology of induction medium and basal medium, which is applied in the field of osteogenic induction medium and osteogenic differentiation, can solve the problems of limiting the effect and research of a single cytokine, achieve the convenience of autologous materials, induce osteogenic differentiation, and facilitate in vitro Amplified effect

Inactive Publication Date: 2017-02-22
GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, the research on the induction of osteogenic differentiation of GMSCs mainly focuses on the different effects of different cytokines on their biological behavior, but is often limited to the research on the effect of a single cytokine

Method used

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  • Osteogenic induction medium and osteogenic differentiation method
  • Osteogenic induction medium and osteogenic differentiation method
  • Osteogenic induction medium and osteogenic differentiation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0060] Example 1 Primary Isolation, Culture, Subculture and Identification of GMSCs

[0061] 1. Primary isolation and culture of GMSCs

[0062] 1) Source of raw materials: The gingiva removed by clinical orthodontic eruption or impacted tooth extraction is used to remove the gingival tissue surrounding the permanent teeth. Patients are required to have no gingival hyperplasia, inflammation and use of drugs that cause gingival hyperplasia. The excised gingival tissue was quickly immersed in 4°C pre-cooled PBS containing 3 times double antibody.

[0063] Among them, the 3-fold double antibody concentration was 300 μ / mL penicillin and 300 μ / mL streptomycin.

[0064] 2) Rinse: Rinse 3 times with PBS containing 3 times the double antibody, and then soak the gingiva in the serum-free medium of mesenchymal stem cells for 5 minutes;

[0065] Among them, the 3-fold double antibody concentration was 300 μ / mL penicillin and 300 μ / mL streptomycin.

[0066] Wherein, the serum-free medi...

Embodiment 2

[0087] Example 2 GMSCs osteogenic differentiation

[0088] In order to verify the effect of osteogenic differentiation induction of the present invention, set 4 groups of control groups and 3 groups of experimental groups at the same time, carry out alizarin red staining, alkaline phosphatase activity determination and osteogenic gene OCN, Col-I to the experimental group and control group. detection.

[0089] Control group 1: a negative control group, a conventional culture group, the culture medium used is: low-sugar DMEM culture medium containing 10% FBS by volume.

[0090] Control group 2: It is a positive control group, which is a conventional mesenchymal stem cell osteogenic induction formula, and the osteogenic induction liquid used is: 50 μg / L vitamin C, 1×10 -8 mol / L dexamethasone, 10 mol / L sodium β-glycerophosphate, volume fraction 10% FBS, and the balance is low-sugar DMEM culture fluid.

[0091] Control group 3: It is a positive control group, and the osteogenic i...

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Abstract

The invention relates to the field of the biotechnology, in particular to an osteogenic induction medium and an osteogenic differentiation method. The osteogenic induction medium is prepared from vitamin C, dexamethasone, beta-sodium glycerol-phosphate, bone morphogenetic protein-2, a vascular endothelial growth factor and a basic medium. Multiple inducing factors in the osteogenic induction medium are combined to act on GMSCs (gingival mesenchymal cells) and have a synergistic effect, the osteogenic differentiation of MSCs can be effectively induced, and the osteogenic differentiation effect is remarkably superior to that of a conventional induction method.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to an osteogenic induction medium and a method for osteogenic differentiation. Background technique [0002] Periodontal disease is not only the main cause of tooth loss in adults, but also a major oral disease that endangers the health of human oral cavity and even the whole body. Periodontal disease is an infectious disease stimulated by chronic inflammation of periodontal tissue. It is an inflammation caused by microorganisms in dental plaque in a corresponding microenvironment. It damages periodontal tissue, especially the destruction of alveolar bone and periodontal tissue. Loss of tissue, eventually leading to loosening of teeth and resorption of alveolar bone. The goal of periodontal treatment is to treat periodontal inflammation, improve the microenvironment to regenerate new connective tissue attachment and support tissue, and complete the construction of alveolar bone, cement...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/077
CPCC12N5/0654C12N2500/38C12N2501/155C12N2501/165C12N2501/30C12N2501/999C12N2506/1392
Inventor 陈海佳葛啸虎王一飞戚康艺张维敏
Owner GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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