Method for analyzing different miRNAs (micro-ribonucleic acids) expression profiles of dental pulp stem cells and stem cells from apical papilla by next-generation sequencing technique

A next-generation sequencing technology and technology of dental pulp stem cells, applied in the field of analyzing the expression profiles of differential miRNAs in two kinds of stem cells, can solve problems such as the inability to efficiently maintain stem cell stemness and regulate the differentiation process

Inactive Publication Date: 2017-10-20
福建医科大学附属口腔医院
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Problems solved by technology

If one molecule or even one pathway is simply controlled, it will not be able to efficiently maintain stem cell stemness and regulate the differentiation process

Method used

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  • Method for analyzing different miRNAs (micro-ribonucleic acids) expression profiles of dental pulp stem cells and stem cells from apical papilla by next-generation sequencing technique
  • Method for analyzing different miRNAs (micro-ribonucleic acids) expression profiles of dental pulp stem cells and stem cells from apical papilla by next-generation sequencing technique

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Embodiment 1

[0025] The method for analyzing the expression profiles of miRNAs differentially between dental pulp stem cells and apical papilla stem cells using next-generation sequencing technology, the steps are as follows:

[0026] (1) Cultivate dental pulp stem cells and apical papilla stem cells: select clinically healthy third molars aged 17-20, remove the pulp or apical papilla tissue under aseptic conditions, cut the tissue into pieces, and use type I Collagenase and dispase were digested at 37°C for 30 minutes, then centrifuged and resuspended, and the resuspension was filtered with a cell sieve to obtain a single cell suspension;

[0027] (2) Sorting of dental pulp stem cells and apical papilla stem cells: the single cell suspension obtained in step (1) was digested with trypsin to collect the cells in the logarithmic growth phase, and STRO-1 was used according to the antibody instructions. - PE labeled cells, collected STRO-1 positive cells by immunomagnetic bead sorting, the co...

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Abstract

The invention provides a method for analyzing different miRNAs (micro-ribonucleic acids) expression profiles of dental pulp stem cells and stem cells from apical papilla by a next-generation sequencing technique. The method includes the steps: preparing single-cell suspensions of the dental pulp stem cells and the stem cells from the apical papilla; separating cells of a logarithmic phase from the single-cell suspensions, and performing marking by the aid of STRO-1-PE; separating and collecting STRO-1 positive cells by the aid of immunomagnetic beads to obtain needed dental pulp stem cells and needed stem cells from the apical papilla; performing extraction by the aid of RNA, and detecting the miRNAs expression profiles of the dental pulp stem cells and the stem cells from the apical papilla by the aid of the next-generation sequencing technique. According to the method, the miRNAs expression profiles of the stem cells are analyzed by the aid of high-throughput next-generation sequencing technique, and the method has profound significance for detecting the different miRNAs expression profiles in the dental pulp stem cells and the stem cells from the apical papilla and knowing characteristics of the stem cells.

Description

technical field [0001] The invention relates to a method for analyzing the differential miRNAs expression profile of dental pulp stem cells and apical papilla stem cells, in particular to a method for analyzing the differential miRNAs expression profiles of the two stem cells using next-generation sequencing technology. Background technique [0002] The teeth are tightly combined with the alveolar bone by means of the roots, and the stability of the roots is very important in the stability of the dentition. Common clinical diseases such as periodontitis, cracked teeth, tooth trauma and other diseases will lead to the loss, fracture, abnormal development of tooth roots, etc., which will eventually lead to tooth loss. After tooth loss, the study of biological root is of great significance. [0003] At present, the study of biological root in vitro is mainly constructed through the method of tissue engineering. One of the commonly used methods is to construct teeth in scaffol...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6869C12Q1/6888C12Q2600/158C12Q2600/178C12Q2535/122
Inventor 吕红兵林诗晗胡雪刚邱在玲
Owner 福建医科大学附属口腔医院
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