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Real-time quantitative PCR analysis of human dental pulp stem cells

ABSTRACT: Real-time quantitative PCR analysis of human dental pulp stem cells

PROVIDER: PRJNA796528 | ENA |

REPOSITORIES: ENA

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Real-time quantitative PCR analysis of human dental pulp stem cells

Project description:Excised pulp tissue was digested in a solution containing 3 mg/ml collagenase type I (Merck KGaA, Darmstadt, Germany) and 4mg/ml dispase (Merck KGaA, Darmstadt, Germany) for 1 h at 37°C. After digestion, 5 volumes of α-MEM medium containing 10% FCS were added. This solution was centrifuged at 120 × g for 10min, at room temperature. The precipitated material was resuspended in α-MEM medium and filtered through filters with pores of 70 μM. This procedure resulted in a single-cell suspension for in vitro culture. Isolated DPSCs were randomly assigned to the following experimental groups: Group 1–LPS stimulus and laser irradiation; Group 2–LPS stimulus and no laser irradiation; Group 3–Control group, no LPS or laser treatment.

2022-01-13 | GSE193448 | GEO

Characteristic expression of MSX1, MSX2, TBX2, and ENTPD1 in dental pulp cells

Project description:Dental pulp cells (DPCs) are a promising source of transplantable cells in regenerative medicine. However, DPCs have not been fully characterized at the molecular level. The purpose of this study was to distinguish DPCs from various source-derived mesenchymal stem cells, fibroblasts, and other cells by the expression of several DPC-characteristic genes. DPCs were isolated from human pulp tissues by the explant method, or the enzyme digestion method, and maintained with media containing 10% serum or 7.5% platelet-rich plasma. RNA was isolated from the cells and from dental pulp tissue specimens. The mRNA levels were determined by DNA microarray and quantitative real-time PCR analyses. The msh homeobox1 (MSX1), msh homeobox 2 (MSX2), T-box 2 (TBX2), and ectonucleoside triphosphate diphosphohydrolase 1 (ENTPD1) mRNA levels in DPCs were higher than the levels found in the following cells: mesenchymal stem cells, derived from bone marrow, synovium, and adipose tissue; and in cells such as fibroblasts, osteoblasts, adipocytes, and chondrocytes. The enhanced expression in DPCs was consistently observed irrespective of donor age, tooth type, and culture medium. Moreover, these genes were expressed at high levels in dental pulp tissue in vivo. We conclude that this gene set may be useful in the identification and characterization of DPCs in basic studies and pulp cell-based regeneration therapy.

2016-06-20 | GSE66084 | GEO

Next Generation Sequencing Facilitates Quantitative Analysis of Normoxia/Hypoxia Human Dental Pulp Cells Transcriptomes

Project description:We report the application of high-throughput profiling of gene expression in human dental pulp cells under normoxia and hypoxia conditions.

2018-08-03 | GSE118046 | GEO

Gene expression analysis of human permanent dental pulp

Project description:Gene expression analysis data of human permanent dental pulp tissues

2016-11-22 | GSE75644 | GEO

Genome-wide maps of mRNAs and ncRNAs in aging and young dental pulp stem cells

Project description:Purpose: The goals of this study are to compare differentially expressed genes in young dental pulp stem cells to the aging dental pulp stem cells and to predict key regular factors. Methods: mRNA and ncRNA profiles of young and aging dental pulp stem cells were generated by deep sequencing, using BGISEQ-500 platform (BGI-Shenzhen, China). The sequence reads that passed quality filters were analyzed at the transcript isoform level with two methods: Agilent Technologies 2100 system and real-time quantitative PCR (QPCR) (TaqMan Probe). Results:After filtering out adaptor-related and low-quality sequences, 669 million clean reads in mRNA Library were generated. Clean reads was mapped to genome sequence with the mapping ratio from 88.54% to 97.61%. A total of 230487 mRNAs was obtained including 123724 known mRNAs and 106763 novel mRNAs respectively; In addition, we detected 177880 lncRNAs，139743 and 38137 were known and novel. For correlation analysis, the coefficient we obtained were all greater than 0.90, indicating a high biological correlation Conclusions: Our study represents the first detailed analysis of differentially expressed genes in aging and young dental pulp stem cells, with biologic replicates, generated by RNA-seq technology. The optimized data analysis workflows reported here should provide a framework for comparative investigations of expression profiles. Our results show that hsa-miR-6724-5p may be a key node in the aging process of DPSCs, and its target genes was involved in the dopaminergic synapse

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Characterization of human dental pulp cells grown in chemically defined serum-free medium

Project description:Dental pulp cells obtained from several donors proliferated actively in a serum-free medium STK2. The growth rate of dental pulp cells from most donors was higher in the serum-free medium than that in a medium containing 10% serum. DNA microarray analyses showed that gene expression profile of dental pulp cells grown in the serum-free medium was similar to that of cells grown in a medium containing 10% serum. However, several genes related to cell proliferation were up-regulated in dental pulp cells grown in the serum-free medium.

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