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Method for separating and preparing dental pulp stem cells

A technology for stem cells and dental pulp, applied in the field of separation and preparation of dental pulp stem cells, can solve problems such as cell loss, and achieve the effects of promoting cell growth, good stability and obvious bacteriostatic effect.

Active Publication Date: 2017-12-22
吉林省太阳鸟再生医学工程有限责任公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Moreover, cell loss is often caused by the fact that dental pulp mesenchymal stem cells are not easy to adhere to the wall.

Method used

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  • Method for separating and preparing dental pulp stem cells
  • Method for separating and preparing dental pulp stem cells
  • Method for separating and preparing dental pulp stem cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] Example 1 Separation and Preparation of Dental Pulp Stem Cells Treated with 0.5 μg / mL Echinacea Chloride

[0053] 1) Collection of teeth: Donate or preserve publicity for juveniles and children who have lost deciduous teeth, or adults who need plastic surgery and wisdom teeth extraction, collect peripheral blood samples from volunteers for virological testing, and collect healthy volunteers after they are qualified Teeth, after routine treatment, were stored in a transport solution containing 1000-2000U / mL penicillin, 1000-2000μg / mL streptomycin and 0.5μg / mL acanthine chloride, refrigerated at 4°C, transported to the laboratory, and stored in Separation process within 24 hours.

[0054] 2) Pulp cleaning and disinfection: transfer the collected teeth to a clean sterile plate, use sterile ophthalmic tweezers to grasp, sequentially wash with 0.9% normal saline containing 0.5 μg / mL acanthine chloride, 75% medical Alcohol, 0.9% normal saline, PBS buffer solution containing ...

Embodiment 2

[0058] Example 2 Separation and Preparation of Dental Pulp Stem Cells Treated with 5 μg / mL Ephedrine Chloride

[0059] 1) Collection of teeth: Donate or preserve publicity for juveniles and children who have lost deciduous teeth, or adults who need plastic surgery and wisdom teeth extraction, collect peripheral blood samples from volunteers for virological testing, and collect healthy volunteers after they are qualified Teeth, after routine treatment, were stored in a transport solution containing 1000-2000U / mL penicillin, 1000-2000μg / mL streptomycin and 5.0μg / mL acanthine chloride, refrigerated at 4°C, transported to the laboratory, and stored in Separation process within 24 hours.

[0060] 2) Pulp cleaning and disinfection: transfer the collected teeth to a clean sterile plate, use sterile ophthalmic tweezers to grasp, and successively wash with 0.9% normal saline containing 5.0 μg / mL amphetine chloride, 75% medical Alcohol, 0.9% normal saline, PBS buffer solution containin...

Embodiment 3

[0064] Example 3 Separation and Preparation of Dental Pulp Stem Cells Treated with 10 μg / mL Echinacea Chloride

[0065] 1) Collection of teeth: Donate or preserve publicity for juveniles and children who have lost deciduous teeth, or adults who need plastic surgery and wisdom teeth extraction, collect peripheral blood samples from volunteers for virological testing, and collect healthy volunteers after they are qualified Teeth, after routine treatment, were stored in a transport solution containing 1000-2000 U / mL penicillin, 1000-2000 μg / mL streptomycin and 10 μg / mL epispinine chloride, refrigerated at 4°C, transported to the laboratory, and stored at 24 separation within hours.

[0066] 2) Pulp cleaning and disinfection: transfer the collected teeth to a clean sterile plate, use sterile ophthalmic tweezers to grasp, and sequentially wash with 0.9% normal saline containing 10 μg / mL acanthine chloride, 75% medical alcohol , 0.9% physiological saline, PBS buffer solution contai...

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Abstract

The invention especially relates to a method for separating and preparing dental pulp stem cells, belonging to the field of cell culture techniques. According to the invention, a natural compound nitidine chloride is used as a bacteriostatic agent in the preparation process of dental pulp stem cells, and the bacteriostatic agent has high safety, good stability and obvious bacteriostatic effect, and can effectively reduce bacterial, fungal or mycoplasma infections; so the method is superior to conventional methods simply using antibiotics. In the process of culture, a culture dish is coated with hyaluronic acid and gelatin, thereby increasing the number of wall-adhering dental pulp stem cells and promoting cell growth, so the purpose of effective amplification is achieved. Experimental results show that the advantages of large cell quantities and low contamination rate are obtained when the method provided by the invention is used for amplification of dental pulp mesenchymal stem cells; and the surface markers of the obtained dental pulp stem cells meet standards in detection, positively expressed surface markers are no less than 95%, and negatively expressed surface markers are no more than 2%.

Description

technical field [0001] The invention relates to the technical field of cell culture, in particular to a method for separating and preparing dental pulp stem cells. Background technique [0002] Stem cells are a kind of pluripotent cells with self-replication ability. Under certain conditions, they can differentiate into various functional cells, and have the potential function of regenerating various tissues, organs and complete individuals. Stem cells come from many sources, including bone marrow, umbilical cord, cord blood, teeth, fat, skin and hair follicles. [0003] Mesenchymal stem cells (MSCs) are pluripotent stem cells that can form various cell types. It has the characteristics of multi-directional differentiation potential, can support hematopoiesis and promote the implantation of hematopoietic stem cells, regulates immunity, and is easy to operate for isolation and culture, and is attracting more and more attention. With the increasing maturity of mesenchymal st...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775A01N1/02
CPCA01N1/0215A01N1/0226C12N5/0664C12N2500/24C12N2500/30C12N2509/00C12N2533/54C12N2533/80
Inventor 杨晓光鲍永利王雪李玉新石晓川李首一
Owner 吉林省太阳鸟再生医学工程有限责任公司
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