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Adipogenesis induction culture medium and adipogenic differentiation method

A technology for inducing medium and basal medium, applied in the field of adipogenic induction medium and adipogenic differentiation, it can solve the problems of limiting the role of a single cytokine, research and other problems, and achieve convenient material collection, easy in vitro expansion, and avoid immune rejection. effect of reaction

Inactive Publication Date: 2017-02-08
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 adipogenic 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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  • Adipogenesis induction culture medium and adipogenic differentiation method
  • Adipogenesis induction culture medium and adipogenic differentiation method
  • Adipogenesis induction culture medium and adipogenic differentiation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0060] Example 1 Primary isolation, culture, passage and identification of GMSCs

[0061] 1. Primary isolation and culture of GMSCs

[0062] 1) Source of raw materials: The gingival tissue surrounding the permanent teeth is excised from the gingival tissue excised by clinical orthodontic eruption or impacted tooth extraction. 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-chilled PBS containing 3x double antibody.

[0063] Among them, the three-fold double antibody concentration is 300u / mL penicillin and 300u / mL streptomycin.

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

[0065] Among them, the three-fold double antibody concentration is 300u / mL penicillin and 300u / mL streptomycin.

[0066] Among them, the serum-free medium of m...

Embodiment 2

[0087] Example 2 Adipogenic differentiation of GMSCs

[0088] 1. Preparation of Platelet Rich Plasma (PRP)

[0089] The peripheral blood of the same donor as the tooth was collected, and the blood samples were transferred into 15mL centrifuge tubes in the ultra-clean bench, 10mL per tube, 2500rpm / min, and centrifuged for 10min. After centrifugation, the blood samples were divided into a pale yellow supernatant layer, an intermediate white cell layer and a red cell bottom layer. Aspirate all the supernatant, the white cell layer and the upper 1-2mm of the red cell layer, transfer it into a new 15mL centrifuge tube, centrifuge at 2500rpm / min for 5min. After the centrifugation, the supernatant layer and the white blood cell layer were aspirated, transferred to a new 15mL centrifuge tube, and centrifuged at 1200g / min for 5min. After centrifugation, the supernatant was discarded, and the bottom 2.5 mL, namely platelet-rich plasma, was retained.

[0090] 2. PRP activation

[009...

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Abstract

The invention relates to the field of the biotechnology, in particular to an adipogenesis induction culture medium and an adipogenic differentiation method. The adipogenesis induction culture medium comprises 3-isobutyl-1-methylxanthine, insulin, indomethacin, dexamethasone, rosiglitazone, PRP (Platelet Rich Plasma) and a basic culture medium. The invention provides an adipogenic differentiation method which enables various inducing factors to be subjected to combined application to act on mesenchymal stem cells including GMSCs and like, has a synergistic effect and can effectively induce the adipogenic differentiation of the mesenchymal stem cells, and the adipogenic differentiation effect of the adipogenic differentiation method is obviously superior to that of an existing conventional induction method. The GMSCs autologous materials are abundant, are easy in expansion in vitro and can be used for autotransplantation so as to avoid immunological rejection reaction.

Description

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

Claims

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Application Information

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IPC IPC(8): C12N5/077
CPCC12N5/0653C12N2500/40C12N2501/33C12N2501/39C12N2501/999C12N2506/1361
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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