Periodontal ligament stem cell culture medium and culture method thereof

A periodontal ligament stem cell and culture method technology, which is applied to the periodontal ligament stem cell culture medium and the field of its culture, can solve the problems of not meeting the scientific research or clinical needs of periodontal ligament stem cells, poor cell activity, slow proliferation rate, etc. The effect of expansion, reduction of production costs, and enhancement of proliferative capacity

Pending Publication Date: 2021-06-04
深圳市皖吉瑞生物医药科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Although the prior art discloses the use of serum-free medium to cultivate periodontal ligament stem cells, the current serum-free medium for periodontal ligament stem cells has problems such as poor cell activity and slow proliferation rate, which cannot meet the requirements of periodontal ligament stem cells. Therefore, it is particularly important to improve the in vitro proliferation ability of periodontal ligament stem cells

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  • Periodontal ligament stem cell culture medium and culture method thereof
  • Periodontal ligament stem cell culture medium and culture method thereof

Examples

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Effect test

Embodiment 1

[0023] A periodontal ligament stem cell culture medium, which consists of the following components: basal medium DMEM / F12 medium, dyphylline, palmatine, PDE 4 inhibitor, glutathione, 2-mercaptoethanol;

[0024] Based on the final concentration of the medium, the mass concentration of each component is: 4.5 μg / mL of dihydroxyprophylline, 11.0 μg / mL of palmatine, 8.5 ng / mL of PDE 4 inhibitor, and 1.5 μg / mL of glutathione , 2-mercaptoethanol 13.0μg / mL.

[0025] A method for culturing periodontal ligament stem cells, comprising the following steps: taking the P3 periodontal ligament stem cells obtained by subculture and resuscitating them with 1×10 4 cells / mL were inoculated in 12-well culture plates, and 1 mL of culture medium was added to each well, at 37°C, 5% CO 2 During the culture, the culture medium was replaced every two days, and the culture was continued for seven days.

Embodiment 2

[0027] A periodontal ligament stem cell culture medium, which consists of the following components: basal medium DMEM / F12 medium, dyphylline, palmatine, PDE 4 inhibitor, glutathione, 2-mercaptoethanol;

[0028] Based on the final concentration of the medium, the mass concentration of each component is: 2.0 μg / mL of dihydroxyprophylline, 9.8 μg / mL of palmatine, 5.0 ng / mL of PDE 4 inhibitor, and 1.0 μg / mL of glutathione , 2-mercaptoethanol 10.0 μg / mL.

[0029] A method for culturing periodontal ligament stem cells, comprising the following steps: taking the P4 periodontal ligament stem cells obtained by subculture and resuscitating them with 1.5×10 4 cells / mL were inoculated in 12-well culture plates, and 1 mL of culture medium was added to each well, at 37°C, 5% CO 2 During the culture, the culture medium was replaced every two days, and the culture was continued for seven days.

Embodiment 3

[0031] A periodontal ligament stem cell culture medium, which consists of the following components: basal medium DMEM / F12 medium, dyphylline, palmatine, PDE 4 inhibitor, glutathione, 2-mercaptoethanol;

[0032] Based on the final concentration of the medium, the mass concentration of each component is: 8.5 μg / mL of dihydroxyprophylline, 12.0 μg / mL of palmatine, 10.0 ng / mL of PDE 4 inhibitor, and 2.0 μg / mL of glutathione , 2-mercaptoethanol 14.5μg / mL.

[0033] A culture method for periodontal ligament stem cells, comprising the following steps: taking the P5 generation periodontal ligament stem cells obtained by subculture and resuscitating them with 2×10 4 cells / mL were inoculated in 12-well culture plates, and 1 mL of culture medium was added to each well, at 37°C, 5% CO 2 During the culture, the culture medium was replaced every two days, and the culture was continued for seven days.

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Abstract

The invention discloses a periodontal ligament stem cell culture medium. The periodontal ligament stem cell culture medium is prepared from the following components: a basic culture medium, dihydroxypropyl theophylline, palmatine, a PDE 4 inhibitor, glutathione and 2-mercaptoethanol. According to the periodontal ligament stem cell culture medium provided by the invention, the diprophylline, the fibrauretin and the PDE 4 inhibitor are added into the culture medium to achieve a synergistic effect, so that periodontal ligament stem cells are stimulated to split, the multiplication capacity of the periodontal ligament stem cells is effectively improved, a large number of periodontal ligament stem cells can be obtained within a short time, the multiplication speed of the cells is increased, a seed cell source is provided for periodontal tissue engineering, and the culture medium is not added with animal serum, so that the safety of clinical application is improved. The invention also provides a culture method of the periodontal ligament stem cells, the culture medium provided by the invention is adopted, P3-P5 generation cells obtained by subculture are taken as culture objects, rapid amplification of the periodontal ligament stem cells is realized, the method is simple and easy to operate, and the production cost is effectively reduced.

Description

technical field [0001] The invention relates to the field of stem cells, in particular to a medium for periodontal ligament stem cells and a culture method thereof. Background technique [0002] At present, periodontal tissue regeneration is an effective way and the ultimate goal for the treatment of periodontal disease. However, basic treatments such as traditional treatment, scaling, and root planing can only eliminate the cause of the disease, but cannot completely restore the lost periodontal tissue, and have little effect. With the development of tissue engineering, researchers have transplanted the complex of seed cells and scaffolds to the defect site, bringing hope for the regeneration of damaged periodontal tissue. [0003] Since Seo et al. isolated periodontal ligament stem cells (PDLSCs) from periodontal ligament tissue in 2004, the cells have been considered as the preferred seed cells for periodontal tissue engineering. They have group self-renewal ability and ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775
CPCC12N5/0664C12N2500/44C12N2501/73C12N2501/999
Inventor 徐会娟陈道林陈龙剑
Owner 深圳市皖吉瑞生物医药科技有限公司
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