scholarly journals Characterisation of proliferation, differentiation potential, and gene expression among clonal cultures of human dental pulp cells

2019 ◽  
Author(s):  
Tomoko Kobayashi ◽  
Daisuke Torii ◽  
Takanori Iwata ◽  
Yuichi Izumi ◽  
Masanori Nasu ◽  
...  
Keyword(s):  
Gene Expression ◽  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Dental Pulp ◽  
Dental Pulp Stem Cells ◽  
Marker Genes ◽  
Differentiation Potential ◽  
Candidate Marker ◽  
Cell Clones ◽  
Human Dental Pulp

Abstract Background: Mesenchymal stem cells are a highly promising source of cells for regeneration therapy because of their multilineage differentiation potential. However, distinct markers for mesenchymal stem cells are not well-established. To identify new candidate marker genes for multipotent human dental pulp stem cells, we analysed the characteristics and gene expression profiles of cell clones obtained from a single dental pulp specimen.Results: Fifty colony-forming single cell-derived clones were isolated from a single dental pulp specimen. These clones varied in their proliferation abilities and surface marker (STRO-1 and CD146) expression patterns, as well as their odontogenic, adipogenic, and chondrogenic differentiation potentials. Four clones maintained their original differentiation potentials during long-term culture. Gene expression profile analysis of five representative clones identified 1227 genes that were related to multipotency. Ninety of these 1227 genes overlapped with genes reportedly involved in ‘stemness or differentiation’. Based on the predicted locations of expressed protein products and large changes in expression levels, 14 of the 90 genes were selected as candidate dental pulp stem cell markers, particularly in relation to their multipotency characteristics.Conclusions: This characterisation of cell clones obtained from a single specimen of human dental pulp provided information regarding new candidate marker genes for multipotent dental pulp stem cells, which could facilitate efficient analysis or enrichment of multipotent stem cells.

Comparison of two digestion strategies on characteristics and differentiation potential of human dental pulp stem cells

2018 ◽  
Vol 93 ◽  
pp. 74-79 ◽  
Author(s):  
Maziar Ebrahimi Dastgurdi ◽  
Fatemeh Ejeian ◽  
Marzie Nematollahi ◽  
Ahmad Motaghi ◽  
Mohammad Hossein Nasr-Esfahani
Keyword(s):  
Stem Cells ◽  
Dental Pulp ◽  
Dental Pulp Stem Cells ◽  
Differentiation Potential ◽  
Human Dental Pulp

scholarly journals The Effect of Mesoporous Bioactive Glass Nanoparticles/Graphene Oxide Composites on the Differentiation and Mineralization of Human Dental Pulp Stem Cells

Nanomaterials ◽  
2020 ◽  
Vol 10 (4) ◽  
pp. 620 ◽  
Author(s):  
Jae Hwa Ahn ◽  
In-Ryoung Kim ◽  
Yeon Kim ◽  
Dong-Hyun Kim ◽  
Soo-Byung Park ◽  
...  
Keyword(s):  
Stem Cells ◽  
Graphene Oxide ◽  
Bioactive Glass ◽  
Dental Pulp ◽  
Dental Pulp Stem Cells ◽  
Mrna Levels ◽  
Alizarin Red ◽  
Differentiation Potential ◽  
Human Dental Pulp ◽  
Mesoporous Bioactive Glass

The purpose of this study was to investigate the effects of mesoporous bioactive glass nanoparticle (MBN)/graphene oxide (GO) composites on the mineralization ability and differentiation potential of human dental pulp stem cells (hDPSCs). MBN/GO composites were synthesized using the sol-gel method and colloidal processing to enhance the bioactivity and mechanical properties of MBN. Characterization using FESEM, XRD, FTIR, and Raman spectrometry showed that the composites were successfully synthesized. hDPSCs were then cultured directly on the MBN/GO (40:1 and 20:1) composites in vitro. MBN/GO promoted the proliferation and alkaline phosphatase (ALP) activity of hDPSCs. In addition, qRT-PCR showed that MBN/GO regulated the mRNA levels of odontogenic markers (dentin sialophosphoprotein (DSPP), dentine matrix protein 1 (DMP-1), ALP, matrix extracellular phosphoglycoprotein (MEPE), bone morphogenetic protein 2 (BMP-2), and runt-related transcription factor 2 (RUNX-2)). The mRNA levels of DSPP and DMP-1, two odontogenesis-specific markers, were considerably upregulated in hDPSCs in response to growth on the MBN/GO composites. Western blot analysis revealed similar results. Alizarin red S staining was subsequently performed to further investigate MBN/GO-induced mineralization of hDPSCs. It was revealed that MBN/GO composites promote odontogenic differentiation via the Wnt/β-catenin signaling pathway. Collectively, the results of the present study suggest that MBN/GO composites may promote the differentiation of hDPSCs into odontoblast-like cells, and potentially induce dentin formation.

scholarly journals Hexosamine-Induced TGF-βSignaling and Osteogenic Differentiation of Dental Pulp Stem Cells Are Dependent on N-Acetylglucosaminyltransferase V

2015 ◽  
Vol 2015 ◽  
pp. 1-11 ◽  
Author(s):  
Yi-Jane Chen ◽  
Chung-Chen Yao ◽  
Chien-Hsun Huang ◽  
Hao-Hueng Chang ◽  
Tai-Horng Young
Keyword(s):  
Stem Cells ◽  
Osteogenic Differentiation ◽  
Dental Pulp ◽  
Critical Role ◽  
Dental Pulp Stem Cells ◽  
Receptor Type ◽  
Marker Genes ◽  
Type I ◽  
Differentiation Potential ◽  
Matrix Deposition

Glycans of cell surface glycoproteins are involved in the regulation of cell migration, growth, and differentiation. N-acetyl-glucosaminyltransferase V (GnT-V) transfers N-acetyl-d-glucosamine to formβ1,6-branched N-glycans, thus playing a crucial role in the biosynthesis of glycoproteins. This study reveals the distinct expression of GnT-V in STRO-1 and CD-146 double-positive dental pulp stem cells (DPSCs). Furthermore, we investigated three types of hexosamines and their N-acetyl derivatives for possible effects on the osteogenic differentiation potential of DPSCs. Our results showed that exogenous d-glucosamine (GlcN), N-acetyl-d-glucosamine (GlcNAc), d-mannosamine (ManN), and acetyl-d-mannosamine (ManNAc) promoted DPSCs’ early osteogenic differentiation in the absence of osteogenic supplements, but d-galactosamine (GalN) or N-acetyl-galactosamine (GalNAc) did not. Effects include the increased level of TGF-βreceptor type I, activation of TGF-βsignaling, and increased mRNA expression of osteogenic differentiation marker genes. The hexosamine-treated DPSCs showed an increased mineralized matrix deposition in the presence of osteogenic supplements. Moreover, the level of TGF-βreceptor type I and early osteogenic differentiation were abolished in the DPSCs transfected with siRNA for GnT-V knockdown. These results suggest that GnT-V plays a critical role in the hexosamine-induced activation of TGF-βsignaling and subsequent osteogenic differentiation of DPSCs.

scholarly journals Different Concentrations of C5a Affect Human Dental Pulp Mesenchymal Stem Cells Differentiation

2021 ◽  
Author(s):  
Jie Liu ◽  
Xiaoling Wei ◽  
Junlong Hu ◽  
Xiaohan Tan ◽  
Xiaocui Kang ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Dental Pulp ◽  
Inhibitory Effect ◽  
Stem Cell Markers ◽  
High Concentration ◽  
Differentiation Potential ◽  
Human Dental Pulp ◽  
Dentin Sialoprotein

Abstract Background During the process of deep decay, when decay approaches the pulp, an immune response is triggered inside the pulp, which activates the complement cascade. The effect of complement component 5a (C5a) on the differentiation of dental pulp mesenchymal stem cells (DPSCs) is related to dentin reparation. The aim of the present study was to stimulate DPSCs with different concentrations of C5a and evaluate the differentiation of odontoblasts using dentin sialoprotein (DSP). Methods DPSCs were divided into the following six groups: i) Control; ii) DPSCs treated with 50 ng/ml C5a; iii) DPSCs treated with 100 ng/ml C5a; iv) DPSCs treated with 200 ng/ml C5a; v) DPSCs treated with 300 ng/ml C5a; and vi) DPSCs treated with 400 ng/ml C5a. Flow cytometry and multilineage differentiation potential were used to identify DPSCs. Mineralization induction, Real-time PCR and Western blot were conducted to evaluate the differentiation of odontoblast in the 6 groups.Result DPSCs can express mesenchymal stem cell markers, including CD105, CD90, CD73 and, a less common marker, mesenchymal stromal cell antigen-1. In addition, DPSCs can differentiate into adipocytes, neurocytes and osteoblasts. All six groups formed mineralized nodules after 28 days of culture. Reverse transcription-quantitative PCR and western blotting indicated that the high concentration C5a groups expressed higher DSP levels and promoted DPSC differentiation, whereas the low concentration C5a groups displayed an inhibitory effect.Conclusion In this study, the increasing concentration of C5a, which accompanies the immune process in the dental pulp, has demonstrated an enhancing effect on odontoblast differentiation at higher C5a concentrations in vitro.

scholarly journals Isolation of Human Dental Pulp Stem Cells and Their Characteristics Before and After Cryopreservation

2021 ◽  
Vol 31 (1) ◽  
pp. 58-69
Author(s):  
Svitlana Mazur ◽  
◽  
Olena Rogulska ◽  
Olena Revenko ◽  
Nataliya Volkova ◽  
...  
Keyword(s):  
Stem Cells ◽  
Dental Pulp ◽  
Third Molar ◽  
Tooth Germ ◽  
Multipotent Mesenchymal Stromal Cells ◽  
Dental Pulp Stem Cells ◽  
Slow Cooling ◽  
Differentiation Potential ◽  
Before And After ◽  
Human Dental Pulp

Dental pulp stem cells (DPSCs) from human third molar tooth germ (wisdom tooth) were isolated using a collagenasebased enzymatic method, the obtained cells were analyzed as for morphology in monolayer culture, immunophenotype, proliferation and differentiation potential before and after cryopreservation. In this study, we showed that based on morphological features, surface markers profile and differentiation potential, the isolated DPSCs corresponded to multipotent mesenchymal stromal cells. DPSCs cryopreservation by slow cooling (1 °С / min) down to –80°C with subsequent immersion into liquid nitrogen in cryoprotectant-free culture medium led to cell death. Cryopreservation using the same protocol in the presence of 10% dimethyl sulfoxide (DMSO) and 20% serum ensured (82 ± 6)% cell viability; while metabolic and proliferative activity, as well as the ability to differentiate into the osteo- and adipogenic lineages of cryopreserved DPSCs were similar to their non-cryopreserved counterparts.

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