Preparation method of dental pulp stem cells (DPSCs)

Abstract

The invention discloses a preparation method of dental pulp stem cells (DPSCs). The preparation method comprises the following steps: 1) conducting disinfection; 2) taking dental pulp, putting the dental pulp into normal saline (1000UI\ml streptomycin, 1000UI\ml penicillin and normal saline) containing double antibodies for flushing, and cutting the dental pulp into small pieces of 1 mm<3>; 3) conducting tissue digestion: using 0.1-0.5% collagenase I, 0.1% Dispase II and normal saline as digestive juice, sealing a digestive tube opening, and performing oscillation digestion for 15-30 minutes at the constant temperature of 37+ / -5 DEG C and the speed of 145-167 rpm; 4) conducting centrifuging, discarding the supernatant, washing with normal saline, conducting centrifuging and discarding thesupernatant; and (5) conducting culture: performing culture by using a mesenchymal stem cell culture medium (5% of platelet lysate and 95% of basal culture medium) at 37+ / -5 DEG C with 5% of carbon dioxide. The method provided by the invention can effectively obtain a single-cell suspension of DPSCs or soften dental pulp tissue, is more beneficial to climbing-out of the DPSCs, improves the obtaining rate of the DPSCs, shortens the cell culture period and guarantees the cell quality.

Description

technical field [0001] The invention belongs to the technical field of cell biology stem cell culture, and in particular relates to a preparation method of dental pulp stem cells. Background technique [0002] Stem cells (Stem Cells, SC) are a kind of pluripotent cells with self-replication ability, which are primitive and unspecialized cells. Under certain conditions, it can differentiate into a variety of functional cells, and has the potential to regenerate various tissues and organs and the human body. Stem cells come from many sources, including bone marrow, umbilical cord, cord blood, teeth and fat. [0003] Dental pulp stem cells were discovered for the first time in 2000, and Gronthos et al. found that dental pulp stem cells existed in deciduous teeth and wisdom teeth in vitro. This is a type of cell with high proliferative ability, high self-renewal ability, and multiple differentiation abilities. Studies have used the cells to successfully construct dental pulp,...

AI Technical Summary

Problems solved by technology

[0006] At present, for the extraction and culture of dental pulp tissue, the traditional methods are direct culture of tissue blocks and collagenase digestion and culture, but the culture period is too long, the amount of cells is small, and it is easy to cause differentiation and incomplete or excessive digestion. and other shortcomings

[0007] 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 by increasing the amount of antibiotics or increasing the level of antibiotics to reduce the chance of contamination, resulting in the abuse of antibiotics

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