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Hypoxia-enhanced Derivation of iPSCs from Human Dental Pulp Cells

ABSTRACT: Hypoxia enhances the reprogramming efficiency of human dermal fibroblasts to become induced pluripotent stem cells (iPSCs). Because we showed previously that the hypoxia facilitates the isolation and maintenance of human dental pulp cells (DPCs), we examined here whether it promotes the reprogramming of DPCs to become iPSCs. To investigate the effect of oxygen concentration on global gene expression, we compared DPCs cultured for 6 days under hypoxia and normoxia.

ORGANISM(S): Homo sapiens

SUBMITTER: Ken-ichi Tezuka

PROVIDER: E-GEOD-45872 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Hypoxia-enhanced derivation of iPSCs from human dental pulp cells.

Iida K K Takeda-Kawaguchi T T Hada M M Yuriguchi M M Aoki H H Tamaoki N N Hatakeyama D D Kunisada T T Shibata T T Tezuka K K

Journal of dental research 20130820 10

Hypoxia enhances the reprogramming efficiency of human dermal fibroblasts to become induced pluripotent stem cells (iPSCs). Because we showed previously that hypoxia facilitates the isolation and maintenance of human dental pulp cells (DPCs), we examined here whether it promotes the reprogramming of DPCs to become iPSCs. Unlike dermal fibroblasts, early and transient hypoxia (3% O2) induced the transition of DPCs to iPSCs by 3.3- to 5.1-fold compared with normoxia (21% O2). The resulting iPSCs c ...[more]

PMID: 23962749

Similar Datasets

Hypoxia-enhanced Derivation of iPSCs from Human Dental Pulp Cells

Project description:Hypoxia enhances the reprogramming efficiency of human dermal fibroblasts to become induced pluripotent stem cells (iPSCs). Because we showed previously that the hypoxia facilitates the isolation and maintenance of human dental pulp cells (DPCs), we examined here whether it promotes the reprogramming of DPCs to become iPSCs.

2013-09-03 | GSE45872 | GEO

miRNA-720 regulates stem cell phenotype, proliferation and differentiation of dental pulp cells

Project description:Small non-coding microRNAs (miRNAs) are known to play crucial roles in stem cell biology, related to cell reprogramming, maintenance of stemness and regulation of cell differentiation. In an attempt to search for novel miRNAs that can control the fate of dental tissue-derived stem cells, we sorted side population (SP) cells, known to be enriched in stem cells, from dental pulp cells (DPCs) and periodontal ligament cells (PDLCs), and performed a miRNA array. As a result, miR-200b and miR-607 were highly expressed in SP cells, whereas miR-1260b and miR-720 was highly expressed in the differentiated main population (MP) cells. Of note, gain-and-loss of function analysis showed that miR-720 regulates the expression of pluripotency factor NANOG and DNA methyltransferases DNMT3A, DNMT3B and DNMT1 in DPCs. Knockdown of miR-720 significantly increased the levels of NANOG as well as the number of cells positive to the stem cell marker SSEA-4, but down-regulated the levels of DNMTs. miR-720 also regulated the proliferation of DPCs as determined by immunocytochemical analysis against ki-67, as well as odontogenic differentiation demonstrated by alizarin red staining, alkaline phosphatase activity and osteopontin mRNA level. Our findings identify mi-720 as a novel miRNA involved in the regulation of stem cell fate of DPCs. Two cell types (dental pulp cells(DPCs), periodontal ligament cells(PDLCs)) were sorted by FACS to isolate side population (SP) and main population (MP) cells. Then a microRNA array was performed with the SP and MP cells of DPCs and PDLCs.

2013-05-17 | E-GEOD-47025 | biostudies-arrayexpress

SAHA Rat Dental Pulp

Project description:Transcriptional response of rat dental pulp cells (DPCs) cultured with SAHA at early and late mineralisation time points Transcript profiling of DPC identified several novel genes expression induced and supressed by HDACi at 24 hrs and 14 days under mineralising conditions. SAHA induces several members of the MMP family of endopepsidases (TIMP-1, MMP-9, MMP-13) and other members of the endochondral ossification pathway at 24 h. 8 experiemental parameters were analysed, each carried out in quadruplicate

2015-06-16 | E-GEOD-67175 | biostudies-arrayexpress

Transcription profiles of human dermal sheath cells cultured under 20%-O2 or 6%-O2 condition

Project description:We investigated the effect of low oxygen culture on the proliferation and hair inductive capacity of human dermal papilla cells (DPCs) and dermal sheath cells (DSCs). DPCs and DSCs were cultured in atmospheric/hyperoxia (20% O2), physiological/normoxia (6% O2), or hypoxia (1% O2) conditions, respectively. Proliferation of DPCs and DSCs was highest under normoxia. Hypoxia inhibited proliferation of DPCs but enhanced proliferation of DSCs. In DPCs, hypoxia down-regulated expression of hair inductive capacity-related genes, including BMP4, LEF1, SOX2, and VCAN, and normoxia up-regulated expression of ALP. In DSCs, both normoxia and hypoxia up-regulated SOX2 expression, and hypoxia down-regulated BMP4 expression. Microarray analysis revealed increased expression of pluripotency-related genes, including SPRY, NR0B1, MSX2, IFITM1, and DAZL, under hypoxia. In an in vivo hair follicle reconstitution assay, cultured DPCs and DSCs were transplanted with newborn mouse epidermal keratinocytes into nude mice using a chamber method. In DPCs, normoxia allowed the most efficient induction of hair follicles. In DSCs, hypoxia allowed the most efficient induction and maturation of hair follicles. These results suggest that low oxygen culture enhances the proliferation and maintains functions of human DPCs and DSCs and could be used for skin engineering and clinical applications.

2021-01-21 | GSE125402 | GEO

Transcription profiles of human dermal papilla cells cultured under 20%-O2 or 6%-O2 condition

2021-01-21 | GSE125401 | 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

Concurrent progress of reprogramming and gene-correction to overcome limitation of gene therapy of ALK2 mutant iPSCs

Project description:To ameliorate the cumbersome processes of reprogramming and subsequent gene-editing in vulnerable iPSCs, we have developed a greatly simplified one-step procedure, simultaneously introducing reprogramming and gene-editing components into human fibroblast cells. This not only serves to save time, labor, and costs, but opens up a new arena of research that is beyond the current application of gene-editing methodologies due to restrictive reprogramming concerns, inhibitory pluripotency maintenance requirements, and vulnerability of single-cell dissociated iPSCs. We generated iPSCs (mALK2-iPSCs) derived from Fibrodysplasia ossificans progressiva (FOP)-fibroblast cells carrying the ACVR1 p.R206H mutation and gene-corrected ALK2-iPSCs (cALK2-iPSCs). To identify gene-correction effects on global gene expression, gene expression profiling was measured in cALK2-iPSCs and mALK2-iPSCs, calibrated to WT-iPSCs with duplication.

2015-05-12 | E-GEOD-68766 | biostudies-arrayexpress

An Automated, High Platform for Induced Pluripotent Stem Cell Derivation, Characterization and Differentiation

Project description:Induced pluripotent stem cells (iPSCs) have become an essential tool for both modeling how causal genetic variants impact cellular function in disease, as well as being an emerging source of tissue for transplantation medicine. Unfortunately the preparation of somatic cells, their reprogramming and the subsequent verification of iPSC pluripotency are laborious, manual processes that limit the scale and level of reproducibility of this technology. Here we describe a modular, robotic platform for iPSC reprogramming that enables automated, high-throughput conversion of skin biopsies into iPSCs and differentiated cells with minimal manual intervention. Using this platform, we demonstrate that automated reprogramming and the pooled selection of pluripotent cells results in high quality, stable, iPSCs. These lines display less line-to-line variation than either manually produced lines or lines produced through automation followed by single colony-subcloning. The robotic platform we describe will enable the application of iPSCs to population-scale biomedical problems including the study of complex genetic diseases and the development of personalized medicines. Two independent human fibroblast lines were reprogrammed, using modified mRNA, into induced pluripotent stem cells (iPSCs). The genomic stability of several cell lines was evaluated using SNP arrays. Three iPSCs originating from one fibroblast line were tested at passages 8 and 20, with two of these derived as picked (clonal) lines and the third being a pooled population. Five iPSCs originating from a second fibroblast line were tested at passages 8 and 20, with three of these derives derived as picked (clonal) lines and two derived as pooled populations. The iPSCs were compared against the original parental fibroblasts.

2015-08-03 | E-GEOD-69868 | biostudies-arrayexpress

SAHA Rat Dental Pulp

2015-06-16 | GSE67175 | GEO

Transcription profiling of human dental pulp stem cells in long term culture

Project description:In previous studies, human dental pulp stem cells (hDPSCs) were mainly isolated from adults. In this manuscript, we tried characterization of hDPSCs isolated from an earlier developmental stage to evaluate potential usage of these cells for tissue regenerative therapy. hDPSCs isolated at the crown-completed stage showed a higher proliferation rate than those isolated at the later stage. When the cells from either group were cultured in medium promoting differentiation towards cells of the osteo/odontoblastic lineage, both became alkaline phosphatase positive, produced calcified matrix, and were also capable of forming dentin-like matrix on scaffolds in vivo. However, during long-term passage, these cells underwent a change in morphology and lost their differentiation ability. The results of a DNA array experiment showed that the expression of a number of genes, such as WNT16, was markedly changed with increasing number of passages, which might have caused the loss of their characteristics as hDPSCs. Experiment Overall Design: To identify genes modulated by passage, we performed cDNA microarray analysis on 3 independent samples at the crown-completed stage (DP2, 28, and 31) that had undergone P4 or P10. Human gene expression was examined by using a Human Genome U133 Plus 2.0 probe array (GeneChip; Affymetrix, Santa Clara, CA, USA), and a Hewlett-Packard Gene Array Scanner (Palo Alto, CA, USA). Each sample was analyzed once. The fluorescence intensity of each probe was quantified by GeneChip Analysis Suite 5.0 (Affymetrix). The level of gene expression was determined as the average difference, obtained by using the GeneChip software.

2008-04-08 | E-GEOD-10444 | biostudies-arrayexpress

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