p16/p53 expression and telomerase activity in immortalized human dental pulp cells

Egbuniwe, Obi; Idowu, Bernadine; Funes, Juan M.; Grant, Andrew; Renton, Tara; Silvio, L. Di

doi:10.4161/cc.10.22.18093

Cited by 32 publications

(30 citation statements)

References 46 publications

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“…It has previously been shown that DPSC can be immortalized using a viral vector containing human telomerase reverse transcriptase ( hTERT ) [10]. We wanted to validate these results and then further investigate morphological, electrophysiological and gene expression changes in immortalized vs. non-immortalized DPSC and DPSC derived neurons.…”

Section: Resultsmentioning

confidence: 98%

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Characterization of neurons from immortalized dental pulp stem cells for the study of neurogenetic disorders

et al. 2015

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A major challenge to the study and treatment of neurogenetic syndromes is accessing live neurons for study from affected individuals. Although several sources of stem cells are currently available, acquiring these involve invasive procedures, may be difficult or expensive to generate and are limited in number. Dental pulp stem cells (DPSC) are multipotent stem cells that reside deep the pulp of shed teeth. To investigate the characteristics of DPSC that make them a valuable resource for translational research, we performed a set of viability, senescence, immortalization and gene expression studies on control DPSC and derived neurons. We investigated the basic transport conditions and maximum passage number for primary DPSC. We immortalized control DPSC using human telomerase reverse transcriptase (hTERT) and evaluated neuronal differentiation potential and global gene expression changes by RNA-seq. We show that neurons from immortalized DPSC share morphological and electrophysiological properties with non-immortalized DPSC. We also show that differentiation of DPSC into neurons significantly alters gene expression for 1305 transcripts. Here we show that these changes in gene expression are concurrent with changes in protein levels of the transcriptional repressor REST/NSRF, which is known to be involved in neuronal differentiation. Immortalization significantly altered the expression of 183 genes after neuronal differentiation, 94 of which also changed during differentiation. Our studies indicate that viable DPSC can be obtained from teeth stored for ≥72hrs, these can then be immortalized and still produce functional neurons for in vitro studies, but that constitutive hTERT immortalization is not be the best approach for long term use of patient derived DPSC for the study of disease.

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Section: Resultsmentioning

confidence: 98%

“…Immortalization was performed as described previously [10]. 1×10 4 cells were plated in a poly-D-lysine coated 12-well dish and infected with pBABE- hTERT - Puro R retrovirus at a multiplicity of infection of 20 plus 6 μg/mL polybrene for 24 hr.…”

Section: Methodsmentioning

confidence: 99%

Characterization of neurons from immortalized dental pulp stem cells for the study of neurogenetic disorders

et al. 2015

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“…The cells were cultured in high‐glucose Dulbecco's modified Eagle medium (DMEM) supplemented with 10% fetal calf serum (FCS), 5 × 1000 U mL −1 penicillin, 5 mg mL −1 streptomycin and 0.11 g L −1 sodium pyruvate (Sigma‐Aldrich). Batches of cultured cells were transfected with retroviral vectors expressing cloned hTERT gene as previously described …”

Section: Methodsmentioning

confidence: 99%

Phenotype-Independent Effects of Retroviral Transduction in Human Dental Pulp Stem Cells

et al. 2013

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An immortalized human dental pulp stem cell (DPSC) line of an odontoblastic phenotype is established to circumvent the normal programmed senescence and to maintain the cell line's usefulness as a tool for further study of cellular activity. DPSCs are isolated from human dental pulp tissues and transfected using hTERT. The influence of this process on the DPSC phenotype and the mRNA expression of oncogenes involved in cellular senescence is investigated. The results reveal an absence of altered DPSC morphology and phenotype following the exogenous introduction of the hTERT gene, which is coupled with a significant reduction in p16 mRNA expression. This provides insight into how to circumvent in vitro dental pulp stem cell death following the exogenous introduction of hTERT.

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“…Ectomesenchymal stem cells (EMSCs), which originate from the cranial neural crest (CNC), are considered to be the progenitors of almost all tooth tissues, except for enamel and maxillofacial mineralized tissues . After migrating to the maxillary and mandibular processes at an early stage of embryogenesis, CNC cells are defined as EMSCs . Then, the EMSCs settle down and interact with dental epithelium, ultimately differentiating into multiple dental tissue‐derived cells.…”

Section: Introductionmentioning

confidence: 99%

SOST, an LNGFR target, inhibits the osteogenic differentiation of rat ectomesenchymal stem cells

Liu

Zhao

et al. 2017

Cell Proliferation

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p16/p53 expression and telomerase activity in immortalized human dental pulp cells

Cited by 32 publications

References 46 publications

Characterization of neurons from immortalized dental pulp stem cells for the study of neurogenetic disorders

Characterization of neurons from immortalized dental pulp stem cells for the study of neurogenetic disorders

Phenotype-Independent Effects of Retroviral Transduction in Human Dental Pulp Stem Cells

SOST, an LNGFR target, inhibits the osteogenic differentiation of rat ectomesenchymal stem cells

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