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Dental pulp stem cell culture medium and culture method

A technology of dental pulp stem cells and a culturing method, applied in the field of dental pulp stem cell culture medium and culture, can solve the problems of inability to obtain dental pulp stem cells in time, low proliferation rate of dental pulp stem cells, easy pollution of dental pulp stem cells, etc., so as to reduce impurities. The probability of bacterial contamination, the effect of avoiding slow cell growth and shortening the attachment time

Pending Publication Date: 2022-01-28
李保建
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, methods commonly used for the preparation of dental pulp stem cells include enzymatic combined digestion, tissue block culture, and tissue block enzymatic digestion. Although all methods can obtain dental pulp stem cells, the preparation process of dental pulp stem cells is not only Contamination, resulting in lower cell numbers and slower growth
In addition, because the dental pulp stem cells are not easy to adhere to the wall, the cell proliferation rate is low, and the ideal quality and quantity of dental pulp stem cells cannot be obtained in time.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] A pulp stem cell medium, which consists of a basic medium DMEM / F12 medium and the following ingredient added in the base medium: boneglum 7.9 ng / ml, an average particle size of 5-20 μm protein stone fine powder 4.7 ng / ml , Sodium selenate 18 ng / ml, L-glutathione 2.3 ng / ml, thick phenol 2.3 ng / ml, BFGF 22 ng / ml, zinc citrate 6.4 μg / ml, β Sodium glycerol phosphate 8 ng / ml.

[0024] Culture of pulp stem cells, including the following steps:

[0025] (1) The pulp stem cells separated from the pulp tissue from the pulp were inoculated in a 12-well culture plate, and the medium was added 1 ml per well, and the concentration of pulp stem cells in the medium was 2 × 10 in the culture medium. 4 / Ml, at 37 ° C, 5% CO 2 In the incubator for primary culture, when the cell fusion reached 75%, 0.25% trypsin was digested and centrifuged after centrifugation;

[0026] (2) Take step (1) Classification Cultured cells inoculated in T75 culture flask, incubation into medium ...

Embodiment 2

[0028] A pulp stem cell medium, composed of base medium DMEM / F12 medium and the following ingredient added in the base medium: boneglum 7.3 ng / ml, the average particle size is 5-20 μm protein stone fine powder 4.5 ng / ml , Sodium selenate 15 ng / ml, L-glutathione 1.5 μg / ml, lactamide 2.8 ng / ml, thick phenol, 20 ng / ml, zinc citrate 5.4 μg / ml, β Sodium glycerol phosphate 5 ng / ml.

[0029] Culture of pulp stem cells, including the following steps:

[0030] (1) The dental pulp stem cells separated from the pulp tissue were inoculated in a 12-well culture plate, and 1 ml of medium was added per well, and the concentration of pulp stem cells in the medium was 4 × 10 in the culture medium. 4 / Ml, at 37 ° C, 5% CO 2 In the incubator, primary culture was carried out. When the cell fusion reached 80%, it was digested with 0.25% trypsin and centrifugation was carried out after centrifugation;

[0031] (2) Taking steps (1) Classification Cultured cells inoculated in T75 cultu...

Embodiment 3

[0033] A pulp stem cell culture medium, which consists of the following ingredients added in the base medium DMEM / F12 medium and the addition of the bonenuts: bonenugose 8.6 ng / ml, with an average particle size of 5-20 μm opal, fine powder 6.1 ng / ml , Sodium selenate 20 ng / ml, L-glutathione 3.2 ng / ml, glutamide 2.7 ng / ml, BFG 25 ng / ml, zinc citrate 6.9μg / ml, β Sodium glycerol phosphate 10 ng / ml.

[0034] Culture of pulp stem cells, including the following steps:

[0035] (1) The pulp stem cells separated from the pulp tissue were inoculated in a 12-well culture plate, and the medium was added 1 ml per well, and the concentration of pulp stem cells in the medium was 5 × 10 in the medium. 4 / Ml, at 37 ° C, 5% CO 2 In the incubator, primary culture was carried out. When the cell fusion reached 85%, it was digested with 0.25% trypsin and centrifugation was centrifuged.

[0036] (2) Taking step (1) Classification Cultured cells inoculated in T75 culture flasks, cultu...

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Abstract

The dental pulp stem cell culture medium comprises a basic culture medium and the following components added in the basic culture medium: ajuga decumbens sugar, opal micro powder, sodium selenite, L-glutathione, piperlongumine, magnolol, bFGF, zinc citrate and beta-sodium glycerophosphate. According to the culture medium provided by the invention, the decumbent bugle herb sugar and the opal micro powder are added; on one hand, the decumbent bugle herb sugar contributes to uniform distribution of the opal micro powder; on the other hand, with the opal micro powder, the adherence of the dental pulp stem cells is improved, the adherence time of the cells is shortened, and the proliferation efficiency of the dental pulp stem cells is improved. Piperlongumine and magnolol are added, so that the probability that the dental pulp stem cells are polluted by infectious microbes in the culture process is reduced, cell apoptosis and slow cell growth caused by the infectious microbes are avoided, and the service time of the culture medium can be prolonged. The invention further provides a culture method of the dental pulp stem cells, the culture medium is adopted, components are safe and clear, and the proliferation efficiency of the dental pulp stem cells is effectively improved.

Description

Technical field [0001] The present invention relates to the field of stem cells, and more particularly to a pulp stem cell culture medium and a culture method. Background technique [0002] Stem cells have multi-directional differentiation potential, which can support hematopoietic and promote hematopoietic stem cell implantation, regulatory immunity and separation and development, and is increasingly concerned. There are many sources of stem cells, including teeth, bone marrow, umbilical cords, cord blood, fat, etc. [0003] The pulp tissue is located within the pulp chamber of the interior of the teeth, and is the only soft tissue in the tooth tissue. In 2000, GRONTHOS et al. Found a cell that has an extremely similar immunotype with bone marrow mesenchymal stem cells and forming mineralized stem cells, and the morphology in cells, can be self-updated And multi-directional differentiation has a strong cloning ability. These fibrous cells separated from the pulp tissue are calle...

Claims

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Application Information

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IPC IPC(8): C12N5/0775
CPCC12N5/0664C12N2501/115C12N2500/34C12N2500/12C12N2500/30C12N2500/42C12N2500/05
Inventor 李保建邱彬
Owner 李保建
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