Preparation method of deciduous tooth mesenchymal stem cells and used kit

A technology of stem cells and kits, applied in the field of stem cell culture, can solve problems such as slow progress in scientific research, achieve the effects of reducing the risk of pathogen infection, convenient use, and no serum

Inactive Publication Date: 2016-08-31
ANHUI NEW LIFE STEM CELL TECH CO LTD
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  • Abstract
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  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, it is urgent to develop a simple, fast and safe method for obtaining a large amount of deciduous tooth mesenchymal stem cells. However, the

Method used

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Embodiment Construction

[0025] The present invention will be further described below in conjunction with specific examples.

[0026] Preparation of kits for preparing primary tooth mesenchymal stem cells:

[0027] 1. Solution configuration:

[0028] (1) Preparation method of deciduous tooth preservation solution: prepare 1L isolated tooth preservation solution, take 0.5L of MEM solution, add penicillin 0.5g, streptomycin 0.8g, metronidazole 0.03g, amphotericin B 0.02g, oxymetholone Base cellulose 8g, insulin 80ug, stir to dissolve, add MEM to 1L constant volume, stir evenly, filter and sterilize with 0.22um filter membrane, aseptically dispense the prepared solution into sterilized collection bottles, each bottle 25ml.

[0029] (2) Preparation method of deciduous tooth lotion: medical saline, 75% alcohol.

[0030] (3) Preparation method of dental pulp digestion solution: weigh and mix according to the proportion that each 1ml of dental pulp digestion solution contains 1-3mg collagenase, 2-4mg DISP...

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Abstract

The invention discloses a preparation method of deciduous tooth mesenchymal stem cells, comprising the steps of: cleaning and sterilizing the tissue surfaces of deciduous teeth with normal saline and 75% alcohol, preserving the deciduous teeth in deciduous teeth preserving fluid; acquiring dental pulp from the deciduous teeth, adding dental pulp digestive fluid to the dental pulp for digestion; obtaining a unicell suspension; adding cell culture fluid, carrying out centrifuging and cleaning, adding precipitated cells into cell culture fluid, putting the cell culture fluid in a carbon dioxide incubator for culturing; when the primary cell fusion reaches 70%, adding cell washing fluid, shaking the culture bottle for washing cells, sucking and abandoning the cell washing fluid, adding cell digestive fluid for digestion, adding cell culture fluid to terminate digestion, repeatedly beating upon the bottle bottom until the cells completely fall off, adding cell culture fluid, inoculating into a culture bottle, putting the culture bottle in the carbon dioxide incubator for culturing, regarding the cells as P1 generation mesenchymal stem cells; when the P1 generation cell fusion reaches 70-80%, carrying out trypsin digestion, collecting digestive cells, carrying out centrifuging and inoculation; after 3d, carrying out trypsin digestion again, collecting digestive cells, carrying out centrifuging, counting and inoculation, and when the P3 generation cell growth reaches 80%, gathering and cryopreserving the cells.

Description

technical field [0001] The present invention relates to the field of stem cell culture, in particular to a method for preparing deciduous tooth mesenchymal stem cells and a kit used in conjunction with them. Background technique [0002] Mesenchymal stem cells (MSCs) are a kind of adult stem cells, which have the characteristics of multi-directional differentiation potential, hematopoietic support and promotion of stem cell implantation, immune regulation and self-replication, etc. Under certain conditions, they can differentiate into Cells derived from various mesoderms such as osteoblast, cartilage, adipose, and myogenic. At present, stem cells are more and more widely used clinically, mainly involving cell transplantation therapy, gene therapy, biological tissue engineering and immunotherapy. [0003] Mesenchymal stem cells are mainly derived from bone marrow, umbilical cord, cord blood, fat, periodontium, etc. Because the acquisition of bone marrow and fat will bring p...

Claims

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Application Information

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IPC IPC(8): C12N5/0775A01N1/02
CPCC12N5/0664A01N1/0221C12N2500/38C12N2501/11C12N2501/115C12N2501/135C12N2501/37C12N2501/39
Inventor 晁若瑜庄莎莎王颖
Owner ANHUI NEW LIFE STEM CELL TECH CO LTD
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