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Culture medium and method for promoting periodontal ligament mesenchymal stem cells to secrete anti-inflammatory factors

A technology of stem cells and anti-inflammatory factors, applied in the field of stem cells, can solve the problems of complex operation steps, limited sources, unsatisfactory effects, etc., and achieve the effect of reducing costs, simple operation, and good anti-inflammatory effect.

Active Publication Date: 2018-09-04
GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Nowadays, bone marrow mesenchymal stem cells or 3D culture are generally used to promote the secretion of anti-inflammatory factors in periodontal inflammation sites, but these methods have defects such as limited sources, high cost, complicated operation steps, or unsatisfactory results.

Method used

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  • Culture medium and method for promoting periodontal ligament mesenchymal stem cells to secrete anti-inflammatory factors
  • Culture medium and method for promoting periodontal ligament mesenchymal stem cells to secrete anti-inflammatory factors
  • Culture medium and method for promoting periodontal ligament mesenchymal stem cells to secrete anti-inflammatory factors

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Embodiment 1, conditioned medium and preparation method thereof

[0022] A method for preparing a conditional medium for promoting the secretion of anti-inflammatory factors from periodontal ligament mesenchymal stem cells, comprising the following steps:

[0023] S11. Isolation and culture of periodontal ligament mesenchymal stem cells

[0024] The freshly extracted healthy third molars were repeatedly rinsed with PBS liquid, and 1 / 3 of the periodontal ligament tissue in the root was scraped; cut into pieces with scissors, added 2 mg / ml type I collagenase (sigma) solution, and placed in a constant temperature shaking box ( After digestion, add an equal amount of DMEM / F12 culture solution containing 10% FBS (DMEM / F12+10% FBS culture solution, Gibco) to stop the digestion, collect the precipitate by centrifugation, and inoculate in a 6-well plate At the same time, place the cover slip, add DMEM / F12+10% FBS culture solution and place at 37°C, 5% CO 2 Cultivate under con...

Embodiment 2

[0029] Example 2, culture medium for promoting periodontal ligament mesenchymal stem cells to secrete anti-inflammatory factors

[0030] The conditioned medium prepared in Example 1 was used to prepare a culture medium for promoting the secretion of anti-inflammatory factors by periodontal ligament-derived mesenchymal stem cells (hereinafter referred to as the medium for promoting anti-inflammatory factor secretion). In this embodiment, the culture medium for promoting anti-inflammatory factor secretion is DMEM / F12 culture medium containing conditioned medium and 10% FBS, and the final protein concentration of the conditioned medium in the medium for promoting anti-inflammatory factor secretion is 50 μg / ml, abbreviated as : DMEM / F12+10% FBS culture solution containing 50 μg / ml conditioned medium.

Embodiment 3

[0031] Embodiment 3, pro-anti-inflammatory factor secretion culture fluid

[0032] The conditioned medium prepared in Example 1 was used to prepare the pro-anti-inflammatory factor secretion culture medium. In this embodiment, the culture medium for promoting anti-inflammatory factor secretion is DMEM / F12 culture medium containing conditioned medium and 10% FBS, and the final protein concentration of the conditioned medium in the medium for promoting anti-inflammatory factor secretion is 100 μg / ml, abbreviated as : DMEM / F12+10% FBS culture solution containing 100 μg / ml conditioned medium.

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Abstract

The invention discloses a culture solution and a method for prompting parodontium mesenchymal stem cells to secrete anti-inflammatory cytokines. The culture solution comprises a conditional medium with the final albumen concentration of 50-200 micrograms / milliliter and a DMEM / F12 culture solution with 10% of FBS. The conditional medium is a culture solution which is already used for culturing the parodontium mesenchymal stem cells. The method includes adding the P1-P5 generation of parodontium mesenchymal stem cells into the DMEM / F12 culture solution with 10% of FBS, culturing the parodontium mesenchymal stem cells at the temperature of 37 DEG C on the condition of 5% of CO2, and culturing the parodontium mesenchymal stem cells in the culture solution for prompting the parodontium mesenchymal stem cells to secrete the anti-inflammatory cytokines for 12-48 hours after the cell merging degree reaches 80-90%. The conditional medium comprises multiple cytokines secreted by the parodontium mesenchymal stem cells, has no toxic and side effects on the cells, has no introduced exogenous materials, is high in safety and low in cost, has better inflammation resistance effect and can promote massive secretion of the anti-inflammatory cytokines.

Description

technical field [0001] The invention relates to the field of stem cells, in particular to a culture medium and a method for promoting periodontal ligament mesenchymal stem cells to secrete anti-inflammatory factors. Background technique [0002] Periodontal disease is the most common oral infectious disease in humans, and its prevalence is relatively high. It is listed as the sixth major complication of diabetes. Inflammation of periodontal tissue, especially periodontitis, is considered a risk factor for certain systemic diseases . In the existing periodontal disease treatment process, most of them are simply treated with periodontal ligament mesenchymal stem cells or traditional Chinese medicine composition. Periodontal ligament stem cells are stem cells that exist in periodontal tissues. They have the characteristics of mesenchymal stem cells, including multi-lineage differentiation potential, obvious high proliferation activity and clone formation ability. They can diff...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/0775C12P21/00
Inventor 王一飞陈海佳葛啸虎马岩岩张维敏
Owner GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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