Method for preparing dental pulp stem cells

A technology of dental pulp stem cells and mesenchymal stem cells, applied in cell dissociation methods, biochemical equipment and methods, animal cells, etc., can solve the problems of dental pulp stem cells such as easy contamination, antibiotic abuse, and increased antibiotics, to ensure purity, Effects of promoting growth and preventing abuse

Inactive Publication Date: 2019-02-12
山东康华生物医疗科技股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Due to the large number and variety of bacteria and fungi in the human oral cavity, dental pulp stem cells are very easy to contaminate during the preparation process. Usually, the use of antibiotics is increased or the level of antibiotics is increased to reduce the chance of contamination, which leads to the abuse of antibiotics
Moreover, the tissue-adherent method of culturing dental pulp stem cells takes a long time

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Embodiment 1: A method for preparing dental pulp stem cells, comprising the following steps:

[0042] S1. Collection of teeth; the collected teeth must be complete, free from caries, pulpitis and pulp necrosis, etc. After collection, the teeth should be stored and transported in the transport solution, and the collected teeth should be placed in the pre-cooled transport solution. Then put it into a 4°C constant temperature transport box and transport it to the laboratory, and separate within 24 hours.

[0043] The transport fluid includes the following components:

[0044] Antimicrobial peptide 10ug / ml;

[0045] Pig bile salt 0.3mg / ml;

[0046] Acetine Chloride 1ug / ml;

[0047] The solvent is sterile PBS.

[0048] Effectively prevent Gram-positive bacteria, Gram-negative bacteria and fungal contamination, and all three are natural compounds, reducing the abuse of antibiotics.

[0049] The collected teeth were disinfected; the teeth were taken out of the ultra-clean...

Embodiment 2

[0069] Embodiment 2: A method for preparing dental pulp stem cells, comprising the following steps:

[0070] S1. Collection of teeth; the collected teeth must be complete, free from caries, pulpitis and pulp necrosis, etc. After collection, the teeth should be stored and transported in the transport solution, and the collected teeth should be placed in the pre-cooled transport solution. Then put it into a 4°C constant temperature transport box and transport it to the laboratory, and separate within 24 hours.

[0071] The transport fluid includes the following components:

[0072] Antimicrobial peptide 20ug / ml;

[0073] Pig bile salt 0.5mg / ml;

[0074] Acetine Chloride 5ug / ml;

[0075] The solvent is sterile PBS.

[0076] Effectively prevent Gram-positive bacteria, Gram-negative bacteria and fungal contamination, and all three are natural compounds, reducing the abuse of antibiotics.

[0077] The collected teeth were disinfected; the teeth were taken out in the ultra-clean...

Embodiment 3

[0097] Embodiment 3: A method for preparing dental pulp stem cells, comprising the following steps:

[0098] S1. Collection of teeth; the collected teeth must be complete, free from caries, pulpitis and pulp necrosis, etc. After collection, the teeth should be stored and transported in the transport solution, and the collected teeth should be placed in the pre-cooled transport solution. Then put it into a 4°C constant temperature transport box and transport it to the laboratory, and separate within 24 hours.

[0099] The transport fluid includes the following components:

[0100] Antimicrobial peptide 30ug / ml;

[0101] Pig bile salt 0.8mg / ml;

[0102] Acetine Chloride 10ug / ml;

[0103] The solvent is sterile PBS.

[0104] Effectively prevent Gram-positive bacteria, Gram-negative bacteria and fungal contamination, and all three are natural compounds, reducing the abuse of antibiotics.

[0105] The collected teeth were disinfected; the teeth were taken out in the ultra-clea...

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PUM

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Abstract

The invention provides a method for preparing dental pulp stem cells. The method comprises the following steps of S1, collecting teeth; S2, separating the dental pulp stem cells, fixing the teeth, splitting the dental crown, fetching out dental pulp, putting the fetched dental pulp into PBS (phosphate buffer saline) for flushing, then preparing the fetched dental pulp into blocks with size of 1 to3 mm<3>, adding a mixed solution of 0.025% to 0.2% of I-type collagenase and 0.025% to 0.2% of neutral protease, digesting for 10 to 30 min at the temperature of 37 DEG C, adding the PBS, centrifuging at the speed of 1000 to 1500 rpm to remove supernatant, settling, adding a culture medium for moistening, attaching the tissue blocks into a cell culture dish, and putting into a CO2 (carbon dioxide) culture box to culture. By adopting the technical scheme, compared with the prior art, the method has the advantages that the enzyme used by the digestion wall-attaching method is a mixture of the I-type collagenase and the neutral protease, and the functions of the I-type collagenase and the neutral protease are mild, so that the restriction of surrounding tissues to the stem cells is reduced,and the damage to the cells is avoided; the purity of the separated stem cells is guaranteed, and the cell harvesting time is shortened.

Description

technical field [0001] The present invention relates to the field of cell biology. [0002] Specifically, it relates to a method for preparing dental pulp stem cells. Background technique [0003] Dental pulp stem cells (DPSCs) are multipotent adult stem cells with the potential to differentiate into neurons, osteoblasts, adipocytes and chondrocytes. Studies have shown that dental pulp stem cells can be used for the repair and regeneration of dentin, dental pulp, and tooth tissue, and can also be used for the repair of skull, jaw, and facial bone injuries. There are also many researches on the application of dental pulp stem cells in other diseases. For example: dental pulp stem cells can be used to treat diseases such as cerebrovascular accidental injury, spinal cord injury, Parkinson's disease, myocardial infarction, diabetes and immunodeficiency. [0004] Due to the large number and variety of bacteria and fungi in the human oral cavity, dental pulp stem cells are extre...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775
CPCC12N5/0664C12N2500/84C12N2509/00C12N2533/54
Inventor 杨致亭任婧婧尚永明王好玉韩伟潘德芹
Owner 山东康华生物医疗科技股份有限公司
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