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A kind of periodontal ligament stem cell cryopreservation solution and cryopreservation method

A technology of periodontal ligament stem cells and cryopreservation method, which is applied in the field of periodontal ligament stem cell cryopreservation and cryopreservation, can solve the problems of periodontal ligament stem cell damage, poor cryopreservation effect, lowering of cell freezing point, etc., so as to achieve prolonged preservation. Time, reduce the cost of cryopreservation, and improve the effect of permeability

Active Publication Date: 2022-07-29
浙江格蕾斯生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

DMSO has a strong ability to penetrate cells and lower the freezing point of cells, thereby reducing the formation of ice crystals and damage to periodontal ligament stem cells. However, DMSO is toxic and interacts with protein hydrophobic groups, which will lead to protein denaturation, thereby Damage the cryopreserved periodontal ligament stem cells, and even adversely affect the survival rate of periodontal ligament stem cells, resulting in poor cryopreservation effect
[0005] Patent CN107114357A discloses a DMSO-free periodontal ligament stem cell cryopreservation solution, but human albumin is added to the cryopreservation solution, and human albumin is a human blood product. Because the raw material comes from human blood, although the raw material plasma Screening of relevant pathogens has been carried out, and measures to remove and inactivate viruses have been added to the production process, but there are still potential risks of spreading some known and unknown pathogens, and clinical use should be more cautious

Method used

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  • A kind of periodontal ligament stem cell cryopreservation solution and cryopreservation method
  • A kind of periodontal ligament stem cell cryopreservation solution and cryopreservation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] A periodontal ligament stem cell cryopreservation solution is composed of the following raw materials by weight: 85 parts of DMEM / F12 medium, 4 parts of beta-sitosterol, 1.2 parts of cinnamaldehyde, 1 part of dilauryl thiodipropionate, 0.7 parts of bakuchiol and 3 parts of epidermal growth factor.

[0029] A method for freezing periodontal ligament stem cells, comprising the following steps:

[0030] (1) Preparation of cryopreservation solution: add β-sitosterol, cinnamaldehyde, dilauryl thiodipropionate, bakuchiol, and epidermal cell growth factor to DMEM / F12 medium, mix well, and store in a 4℃ for standby;

[0031] (2) Preparation of periodontal ligament stem cell suspension: P5 generation periodontal ligament stem cells were added to the cryopreserved solution prepared in step (1) and mixed to obtain a cell suspension, and the cell density in the suspension was 1×10 6 cells / mL, aliquot into sterile cryopreservation tubes, and add 1.5 mL of cell suspension to each t...

Embodiment 2

[0034] A periodontal ligament stem cell cryopreservation solution is composed of the following raw materials in parts by weight: 80 parts of DMEM / F12 medium, 2.5 parts of beta-sitosterol, 0.9 part of cinnamaldehyde, 0.5 part of dilauryl thiodipropionate, 0.5 part of bakuchiol and 1 part of epidermal growth factor.

[0035] A method for freezing periodontal ligament stem cells, comprising the following steps:

[0036] (1) Preparation of cryopreservation solution: add β-sitosterol, cinnamaldehyde, dilauryl thiodipropionate, bakuchiol, and epidermal cell growth factor to DMEM / F12 medium, mix well, and store in a 4℃ for standby;

[0037] (2) Preparation of periodontal ligament stem cell suspension: P5 generation periodontal ligament stem cells were added to the cryopreserved solution prepared in step (1) and mixed to obtain a cell suspension, and the cell density in the suspension was 3×10 6 cells / mL, aliquot into sterile cryopreservation tubes, and add 1.5 mL of cell suspension...

Embodiment 3

[0040] A periodontal ligament stem cell cryopreservation solution is composed of the following raw materials in parts by weight: 90 parts of DMEM / F12 medium, 5 parts of beta-sitosterol, 1.5 parts of cinnamaldehyde, 1.2 parts of dilauryl thiodipropionate, 0.8 parts of bakuchiol and 5 parts of epidermal growth factor.

[0041] A method for freezing periodontal ligament stem cells, comprising the following steps:

[0042] (1) Preparation of cryopreservation solution: add β-sitosterol, cinnamaldehyde, dilauryl thiodipropionate, bakuchiol, and epidermal cell growth factor to DMEM / F12 medium, mix well, and store in a 4℃ for standby;

[0043] (2) Preparation of periodontal ligament stem cell suspension: P5 generation periodontal ligament stem cells were added to the cryopreserved solution prepared in step (1) and mixed to obtain a cell suspension, and the cell density in the suspension was 5×10 6 cells / mL, aliquot into sterile cryopreservation tubes, and add 1.5 mL of cell suspensi...

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Abstract

The invention discloses a periodontal ligament stem cell cryopreservation solution, which is composed of the following raw materials in parts by weight: 80-90 parts of DMEM / F12 medium, 2.5-5 parts of beta-sitosterol, 0.9-1.5 parts of cinnamaldehyde, 0.5-1.2 parts of dilauryl dipropionate, 0.5-0.8 parts of bakuchiol, and 1-5 parts of epidermal cell growth factor. The periodontal ligament stem cell cryopreservation solution provided by the invention does not add DMSO and human serum albumin, but adds β-sitosterol and cinnamaldehyde as cryoprotectants to improve the permeability of cell membranes and reduce the formation of ice crystals. Dilauryl thiodipropionate and bakuchiol are also added to maintain the stability of the cryopreservation solution, so that the periodontal ligament stem cells are dispersed and fully infiltrated by the surrounding cryopreservation solution, preventing the cells from precipitation during the cryopreservation process. It accumulates into clusters and is protected during cryopreservation to reduce damage, and the components work synergistically to improve cell survival. The invention also provides a method for freezing periodontal ligament stem cells, which is simple and easy to operate, and effectively reduces the freezing storage cost of periodontal ligament stem cells.

Description

technical field [0001] The invention relates to the field of stem cells, in particular to a periodontal ligament stem cell cryopreservation solution and a cryopreservation method. Background technique [0002] Periodontal ligament stem cells (PDLSCs) are a kind of fibroblast-like stem cells with self-renewal, strong proliferation ability and multi-directional differentiation ability existing in the periodontal tissue of teeth. Periodontal ligament stem cells have the potential of multi-directional differentiation, which can be differentiated into adipocytes, osteoblast-like cells, cementoblast-like cells and neural precursor cells in vitro, and can be differentiated into cementoblast-like and periodontal cells in vivo membranous tissue. Therefore, periodontal ligament stem cells are widely used in clinical practice. [0003] Cryopreservation of periodontal ligament stem cells is an important means to ensure the clinical application of periodontal ligament stem cells. Beca...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01N1/02
CPCA01N1/0221A01N1/021
Inventor 徐会娟陈道林陈龙剑
Owner 浙江格蕾斯生物科技有限公司