scholarly journals piggyBac Transposon-Based Immortalization of Human Deciduous Tooth Dental Pulp Cells with Multipotency and Non-Tumorigenic Potential

2019 ◽  
Vol 20 (19) ◽  
pp. 4904 ◽  
Author(s):  
Emi Inada ◽  
Issei Saitoh ◽  
Naoko Kubota ◽  
Yoko Iwase ◽  
Yuki Kiyokawa ◽  
...  
Keyword(s):  
Cell Lines ◽  
Dental Pulp ◽  
Fluorescent Protein ◽  
Deciduous Tooth ◽  
Specific Factor ◽  
Dental Pulp Cells ◽  
Differentiation Potential ◽  
Normal Medium ◽  
Cdna Expression ◽  
Pulp Cells

We aimed to immortalize primarily isolated human deciduous tooth-derived dental pulp cells (HDDPCs) by transfection with piggyBac (PB)-based transposon vectors carrying E7 from human papilloma virus 16 or complementary DNA (cDNA) encoding human telomerase reverse transcriptase (hTERT). HDDPCs were co-transfected with pTrans (conferring PB transposase expression) + pT-pac (conferring puromycin acetyltransferase expression) + pT-tdTomato (conferring tdTomato cDNA expression) and pT-E7 (conferring E7 expression) or pTrans + pT-pac + pT-EGFP (conferring enhanced green fluorescent protein cDNA expression) + pT-hTERT (conferring hTERT expression). After six days, these cells were selected in medium containing 5 μg/mL puromycin for one day, and then cultured in normal medium allowing cell survival. All resultant colonies were harvested and propagated as a pool. Stemness and tumorigenic properties of the established cell lines (“MT_E7” for E7 and “MT_hTERT” for hTERT) with untransfected parental cells (MT) were examined. Both lines exhibited proliferation similar to that of MT, with alkaline phosphatase activity and stemness-specific factor expression. They displayed differentiation potential into multi-lineage cells with no tumorigenic property. Overall, we successfully obtained HDDPC-derived immortalized cell lines using a PB-based transfection system. The resultant and parental cells were indistinguishable. Thus, E7 and hTERT could immortalize HDDPCs without causing cancer-associated changes or altering phenotypic properties.

scholarly journals Sclerostin promotes human dental pulp cells senescence

PeerJ ◽  
2018 ◽  
Vol 6 ◽  
pp. e5808 ◽  
Author(s):  
Yanjing Ou ◽  
Yi Zhou ◽  
Shanshan Liang ◽  
Yining Wang
Keyword(s):  
Dental Pulp ◽  
Wnt Signaling Pathway ◽  
Dental Pulp Cells ◽  
Differentiation Potential ◽  
Human Dental Pulp Cells ◽  
Qrt Pcr ◽  
Odontoblastic Differentiation ◽  
Human Dental Pulp ◽  
Pulp Cells

Background Senescence-related impairment of proliferation and differentiation limits the use of dental pulp cells for tissue regeneration. Deletion of sclerostin improves the dentinogenesis regeneration, while its role in dental pulp senescence is unclear. We investigated the role of sclerostin in subculture-induced senescence of human dental pulp cells (HDPCs) and in the senescence-related decline of proliferation and odontoblastic differentiation. Methods Immunohistochemical staining and qRT-PCR analyses were performed to examine the expression pattern of sclerostin in young (20–30-year-old) and senescent (45–80-year-old) dental pulps. HDPCs were serially subcultured until senescence, and the expression of sclerostin was examined by qRT-PCR analysis. HDPCs with sclerostin overexpression and knockdown were constructed to investigate the role of sclerostin in HDPCs senescence and senescence-related impairment of odontoblastic differentiation potential. Results By immunohistochemistry and qRT-PCR, we found a significantly increased expression level of sclerostin in senescent human dental pulp compared with that of young human dental pulp. Additionally, elevated sclerostin expression was found in subculture-induced senescent HDPCs in vitro. By sclerostin overexpression and knockdown, we found that sclerostin promoted HDPCs senescence-related decline of proliferation and odontoblastic differentiation potential with increased expression of p16, p53 and p21 and downregulation of the Wnt signaling pathway. Discussion The increased expression of sclerostin is responsible for the decline of proliferation and odontoblastic differentiation potential of HDPCs during cellular senescence. Anti-sclerostin treatment may be beneficial for the maintenance of the proliferation and odontoblastic differentiation potentials of HDPCs.

scholarly journals Highly Proliferative Immortalized Human Dental Pulp Cells Retain the Odontogenic Phenotype when Combined with a Beta-Tricalcium Phosphate Scaffold and BMP2

2020 ◽  
Vol 2020 ◽  
pp. 1-18 ◽  
Author(s):  
Xiangfen Li ◽  
Liu Wang ◽  
Qin Su ◽  
Ling Ye ◽  
Xuedong Zhou ◽  
...  
Keyword(s):  
Tricalcium Phosphate ◽  
Dental Pulp ◽  
Biological Behavior ◽  
Dental Pulp Cells ◽  
Differentiation Potential ◽  
Human Dental Pulp Cells ◽  
Beta Tricalcium Phosphate ◽  
Human Dental Pulp ◽  
Pulp Cells

Human dental pulp cells (HDPCs) play a vital role in dentin formation and reparative dentinogenesis, which indicated their potential application in regenerative medicine. However, HDPCs, which can only be obtained from scarce human pulp tissues, also have a limited lifespan in vitro, and stem cells usually lose their original characteristics over a large number of passages. To overcome these challenges, we successfully immortalized human dental pulp cells using the piggyBac system which was employed to efficiently overexpress the SV40 T-Ag, and we then comprehensively described the cell biological behavior. The immortalized human dental pulp cells (iHDPCs) acquired long-term proliferative activity and expressed most HDPC markers. The iHDPCs maintained multiple differentiation potential and could be induced to differentiate into chondrogenic, osteogenic, and adipogenic cells in vitro. We also proved that the iHDPCs gained a stronger ability to migrate than the primary cells, while apoptosis was inhibited. Furthermore, highly proliferative iHDPCs displayed no oncogenicity when subcutaneously implanted into athymic nude mice. Finally, iHDPCs exhibited odontogenic differentiation ability and secreted dentin sialophosphoprotein (DSPP) when combined with a beta-tricalcium phosphate scaffold and bone morphogenetic protein-2 (BMP2) in vivo. Conclusively, the established iHDPCs are a valuable resource for mechanistic study of dental pulp cell differentiation and dental pulp injury repair, as well as for applications in tooth regeneration.

scholarly journals Choice of Feeders is Important When First Establishing iPSCs Derived from Primarily Cultured Human Deciduous Tooth Dental Pulp Cells

Cell Medicine ◽  
2015 ◽  
Vol 8 (1-2) ◽  
pp. 9-23 ◽  
Author(s):  
Issei Saitoh ◽  
Emi Inada ◽  
Yoko Iwase ◽  
Hirofumi Noguchi ◽  
Tomoya Murakami ◽  
...  
Keyword(s):  
Dental Pulp ◽  
Deciduous Tooth ◽  
Dental Pulp Cells ◽  
Pulp Cells

scholarly journals Time-Dependent Response of Human Deciduous Tooth-Derived Dental Pulp Cells Treated with TheraCal LC: Functional Analysis of Gene Interactions Compared to MTA

2020 ◽  
Vol 9 (2) ◽  
pp. 531 ◽  
Author(s):  
Ok Hyung Nam ◽  
Jae-Hwan Kim ◽  
Sung Chul Choi ◽  
Young Kim
Keyword(s):  
Functional Analysis ◽  
Dental Pulp ◽  
Enrichment Analysis ◽  
Deciduous Tooth ◽  
Functional Enrichment ◽  
Receptor Interaction ◽  
Dental Pulp Cells ◽  
Pulp Tissue ◽  
Pulp Capping ◽  
Pulp Cells

Pulp capping material should facilitate hard tissue regeneration on the injured pulp tissue. TheraCal LC (TC) was recently developed. Although TC has shown reliable clinical outcomes after direct pulp capping, there are still remaining concerns regarding its detrimental effect on pulp cells. Therefore, this study aimed to identify the gene expression of human deciduous tooth-derived dental pulp cells exposed to TC compared to mineral trioxide aggregate (MTA). The cells were cultured and exposed to TC and MTA for 24 and 72 h. Next, total RNA was isolated. QuantSeq 3′ mRNA-sequencing was used to examine differentially expressed genes (DEGs) in exposed to TC and MTA. Functional analysis of DEGs was performed using bioinformatics analysis. In gene ontology (GO) functional enrichment analysis, cells in TC for 24 h presented significantly enriched immune response (p < 0.001) and inflammatory response (p < 0.01) compared to MTA. TC showed enriched positive regulation of cell migration at 72 h (p < 0.001). In Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis, neuroactive ligand–receptor interaction (p = 1.19 × 10−7) and calcium signaling pathway (p = 2.96 × 10−5) were confirmed in the shared DEGs in TC. In conclusion, DEGs in TC may be involved in pathways associated with osteoclastogenesis and osteoclastic differentiation.

scholarly journals Odontogenic differentiation potential of human dental pulp cells cultured on a calcium-aluminate enriched chitosan-collagen scaffold

2017 ◽  
Vol 21 (9) ◽  
pp. 2827-2839 ◽  
Author(s):  
Diana Gabriela Soares ◽  
Hebert Luís Rosseto ◽  
Débora Salles Scheffel ◽  
Fernanda Gonçalves Basso ◽  
Claudia Huck ◽  
...  
Keyword(s):  
Calcium Aluminate ◽  
Dental Pulp ◽  
Collagen Scaffold ◽  
Dental Pulp Cells ◽  
Differentiation Potential ◽  
Human Dental Pulp Cells ◽  
Odontogenic Differentiation ◽  
Human Dental Pulp ◽  
Pulp Cells ◽  
Cells Cultured

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

Human Cell ◽  
2020 ◽  
Vol 33 (3) ◽  
pp. 490-501
Author(s):  
Tomoko Kobayashi ◽  
Daisuke Torii ◽  
Takanori Iwata ◽  
Yuichi Izumi ◽  
Masanori Nasu ◽  
...  
Keyword(s):  
Gene Expression ◽  
Dental Pulp ◽  
Dental Pulp Cells ◽  
Differentiation Potential ◽  
Human Dental Pulp Cells ◽  
Clonal Cultures ◽  
Human Dental Pulp ◽  
Pulp Cells

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

2020 ◽  
Author(s):  
Tomoko Kobayashi ◽  
Daisuke Torii ◽  
Takanori Iwata ◽  
Yuichi Izumi ◽  
Masanori Nasu ◽  
...  
Keyword(s):  
Gene Expression ◽  
Dental Pulp ◽  
Dental Pulp Cells ◽  
Differentiation Potential ◽  
Human Dental Pulp Cells ◽  
Clonal Cultures ◽  
Human Dental Pulp ◽  
Pulp Cells ◽  
Journal Submission

Abstract The authors have withdrawn the journal submission associated with this preprint and requested that the preprint also be withdrawn.

scholarly journals FAM20C could be targeted by TET1 to promote odontoblastic differentiation potential of human dental pulp cells

2017 ◽  
Vol 51 (2) ◽  
pp. e12426 ◽  
Author(s):  
Qimeng Li ◽  
Baicheng Yi ◽  
Zhihui Feng ◽  
Runsha Meng ◽  
Cheng Tian ◽  
...  
Keyword(s):  
Dental Pulp ◽  
Dental Pulp Cells ◽  
Differentiation Potential ◽  
Human Dental Pulp Cells ◽  
Odontoblastic Differentiation ◽  
Human Dental Pulp ◽  
Pulp Cells
Sign in / Sign up

Export Citation Format

Share Document