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Culture medium for human dental pulp stem cells and preparation method for human dental pulp stem cells

A technology of dental pulp stem cells and culture medium is applied in the field of preparation of human dental pulp stem cell culture medium and human dental pulp stem cells, which can solve the problems of rejection, increase the risk of clinical application of dental pulp stem cells, etc. Good for growth and less damage

Inactive Publication Date: 2017-09-19
SHENZHEN WINGOR BIO TECH
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AI Technical Summary

Problems solved by technology

At present, the culture system of dental pulp stem cells mainly adopts the medium containing animal-derived fetal bovine serum. Animal-derived fetal bovine serum may cause immune rejection in humans to a certain extent, which increases the risk of clinical application of dental pulp stem cells.

Method used

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  • Culture medium for human dental pulp stem cells and preparation method for human dental pulp stem cells

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preparation example Construction

[0023] The present invention also provides a preparation method of human dental pulp stem cells based on the medium described in the above technical scheme, comprising the following steps:

[0024] 1) Cleaning and disinfecting tooth raw materials by placing them in 0.9% sodium chloride solution containing penicillin sodium and streptomycin sulfate to obtain sterilized teeth, the tooth raw materials being fallen deciduous teeth or wisdom teeth;

[0025] 2) taking out the pulp tissue from the sterilized teeth described in step 1), placing it in a serum-free medium, and cutting the pulp tissue into tissue pieces;

[0026] 3) taking the tissue block obtained in step 2) and placing it in human dental pulp stem cell culture medium, and covering it with a cover glass;

[0027] 4) On the 4th to 6th day of culture, replace the medium; on the 10th to 12th day of culture, replace half of the medium; on the 15th to 18th day of culture, the cell confluence rate reaches 70 to 80%, and subcu...

Embodiment 1

[0038] Place two 10cm sterile petri dishes in a biological safety cabinet, add 40ml of 0.9% sodium chloride injection containing double antibodies (penicillin sodium (100U / ml), streptomycin sulfate (100mg / ml)). Use sterile tweezers to take out the teeth in the collection bottle, put them into a petri dish containing double antibody and 0.9% sodium chloride injection, clean and disinfect for 2 minutes, fully remove periodontal soft tissue and blood, and put the sterilized teeth into Sterilize again in the second Petri dish.

[0039] Put the cleaned tooth on sterile gauze, then wrap the tooth with gauze, use a bench vise to force open the enamel and dentin of the tooth, open the sterile gauze, and use tweezers to find out the pulp from the pulp cavity Tissues were loaded into EP tubes containing serum-free medium.

[0040] Under the condition of sufficient infiltration of serum-free medium, use sterile ophthalmic scissors to cut the pulp tissue into about 1.0mm 3 The size of t...

Embodiment 2

[0044] Place two 10cm sterile petri dishes in a biological safety cabinet, add 40ml of 0.9% sodium chloride injection containing double antibodies (penicillin sodium (100U / ml), streptomycin sulfate (100mg / ml)). Use sterile tweezers to take out the teeth in the collection bottle, put them into a petri dish containing double antibody and 0.9% sodium chloride injection, clean and disinfect for 3 minutes, fully remove periodontal soft tissue and blood, and put the sterilized teeth into Sterilize again in the second Petri dish.

[0045] Put the cleaned tooth on sterile gauze, then wrap the tooth with gauze, use a bench vise to force open the enamel and dentin of the tooth, open the sterile gauze, and use tweezers to find out the pulp from the pulp cavity Tissues were loaded into EP tubes containing serum-free medium.

[0046] Under the condition of sufficient infiltration of serum-free medium, use sterile ophthalmic scissors to cut the pulp tissue into about 1.0mm 3 The size of t...

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Abstract

The invention relates to a culture medium for human dental pulp stem cells and a preparation method for human dental pulp stem cells and belongs to the technical field of stem cell preparation. The culture medium for human dental pulp stem cells, provided by the invention, is based on an alpha-MEM culture medium and contains 5%-10% of human AB serum by volume percent, 100mu M ascorbic acid, 2mM L-glutamine, 100U / ml penicillin sodium and 100mg / ml streptomycin. The dental pulp stem cells prepared by the culture medium provided by the invention are high in yield, are free from human immunological rejection and can supply reliable seed cells for repairing and regeneration of clinical teeth.

Description

technical field [0001] The invention relates to the technical field of stem cell preparation, in particular to a culture medium for human dental pulp stem cells and a preparation method for human dental pulp stem cells. Background technique [0002] Stem cells are cells with self-replication and multi-directional differentiation capabilities, which can continuously self-renew and differentiate into one or more cells that constitute human tissues or organs under specific conditions, which can restore the tissue's ability to regenerate and restore its functions. Researchers discovered a new type of adult stem cell in the pulp tissue of exfoliated deciduous teeth, and named it deciduous dental pulp stem cells. So far, many experiments have proved that deciduous dental pulp stem cells have a high degree of self-renewal, clone formation and osteogenic differentiation. ability. [0003] Dental pulp stem cells of deciduous teeth have the following advantages: 1) It is convenient t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775
CPCC12N5/0664C12N2500/32C12N2500/38C12N2500/84
Inventor 姜舒张芸纪惜銮杨顺罗朝霞
Owner SHENZHEN WINGOR BIO TECH
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