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Analysis of gene expression during odontogenic differentiation of cultured human dental pulp cells.

Seo MS, Hwang KG, Kim H, Baek SH - Restor Dent Endod (2012)

Bottom Line: Total RNA was extracted from cells and analyzed by microarray analysis and the key results were confirmed selectively by reverse-transcriptase polymerase chain reaction (RT-PCR).GSEA revealed that in the induced cells, genes implicated in Apoptosis and Signaling by wingless MMTV integration (Wnt) were significantly upregulated.Genes implicated in Apoptosis and Signaling by Wnt are highly connected to the differentiation of dental pulp cells into odontoblast.

View Article: PubMed Central - PubMed

Affiliation: Department of Dentistry, Hanyang University College of Medicine, Seoul, Korea. ; Department of Conservative Dentistry, Seoul National University School of Dentistry, Seoul, Korea.

ABSTRACT

Objectives: We analyzed gene-expression profiles after 14 day odontogenic induction of human dental pulp cells (DPCs) using a DNA microarray and sought candidate genes possibly associated with mineralization.

Materials and methods: Induced human dental pulp cells were obtained by culturing DPCs in odontogenic induction medium (OM) for 14 day. Cells exposed to normal culture medium were used as controls. Total RNA was extracted from cells and analyzed by microarray analysis and the key results were confirmed selectively by reverse-transcriptase polymerase chain reaction (RT-PCR). We also performed a gene set enrichment analysis (GSEA) of the microarray data.

Results: Six hundred and five genes among the 47,320 probes on the BeadChip differed by a factor of more than two-fold in the induced cells. Of these, 217 genes were upregulated, and 388 were down-regulated. GSEA revealed that in the induced cells, genes implicated in Apoptosis and Signaling by wingless MMTV integration (Wnt) were significantly upregulated.

Conclusions: Genes implicated in Apoptosis and Signaling by Wnt are highly connected to the differentiation of dental pulp cells into odontoblast.

No MeSH data available.


Related in: MedlinePlus

(a) Left: Dishes of control (non-induced) DPCs, Right: induced (OM-treated) DPCs after Alizarin-Red staining; (b) The expression of surface markers as analyzed by flow cytometry of control and induced DPCs. Left: control DPCs. Right: induced DPCs. Q3 means cells exhibits negative to STRO-1 and CD146 antibody. DPC, dental pulp cell; OM, odontogenic induction medium.
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Figure 1: (a) Left: Dishes of control (non-induced) DPCs, Right: induced (OM-treated) DPCs after Alizarin-Red staining; (b) The expression of surface markers as analyzed by flow cytometry of control and induced DPCs. Left: control DPCs. Right: induced DPCs. Q3 means cells exhibits negative to STRO-1 and CD146 antibody. DPC, dental pulp cell; OM, odontogenic induction medium.

Mentions: To investigate the hard tissue producing potential of human DPCs undergoing odontogenic differentiation in OM, we assessed Alizarin-Red staining, and levels of transcripts of differentiation markers. Mineralization nodule formation was seen in the DPCs incubated in OM for 14 days (Figure 1a). To determine the effects of OM on the stem cell properties of DPCs, we tested cells for the stem cell markers, STRO-1 and CD 146. Flow cytometric analysis revealed that the OM-treated cells expressed very much reduced levels of CD146 (0.03%) and STRO-1 (0.7%) (Figure 1b). This may suggest that the OM-treated DPCs contain a low proportion of stem cells.


Analysis of gene expression during odontogenic differentiation of cultured human dental pulp cells.

Seo MS, Hwang KG, Kim H, Baek SH - Restor Dent Endod (2012)

(a) Left: Dishes of control (non-induced) DPCs, Right: induced (OM-treated) DPCs after Alizarin-Red staining; (b) The expression of surface markers as analyzed by flow cytometry of control and induced DPCs. Left: control DPCs. Right: induced DPCs. Q3 means cells exhibits negative to STRO-1 and CD146 antibody. DPC, dental pulp cell; OM, odontogenic induction medium.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3569398&req=5

Figure 1: (a) Left: Dishes of control (non-induced) DPCs, Right: induced (OM-treated) DPCs after Alizarin-Red staining; (b) The expression of surface markers as analyzed by flow cytometry of control and induced DPCs. Left: control DPCs. Right: induced DPCs. Q3 means cells exhibits negative to STRO-1 and CD146 antibody. DPC, dental pulp cell; OM, odontogenic induction medium.
Mentions: To investigate the hard tissue producing potential of human DPCs undergoing odontogenic differentiation in OM, we assessed Alizarin-Red staining, and levels of transcripts of differentiation markers. Mineralization nodule formation was seen in the DPCs incubated in OM for 14 days (Figure 1a). To determine the effects of OM on the stem cell properties of DPCs, we tested cells for the stem cell markers, STRO-1 and CD 146. Flow cytometric analysis revealed that the OM-treated cells expressed very much reduced levels of CD146 (0.03%) and STRO-1 (0.7%) (Figure 1b). This may suggest that the OM-treated DPCs contain a low proportion of stem cells.

Bottom Line: Total RNA was extracted from cells and analyzed by microarray analysis and the key results were confirmed selectively by reverse-transcriptase polymerase chain reaction (RT-PCR).GSEA revealed that in the induced cells, genes implicated in Apoptosis and Signaling by wingless MMTV integration (Wnt) were significantly upregulated.Genes implicated in Apoptosis and Signaling by Wnt are highly connected to the differentiation of dental pulp cells into odontoblast.

View Article: PubMed Central - PubMed

Affiliation: Department of Dentistry, Hanyang University College of Medicine, Seoul, Korea. ; Department of Conservative Dentistry, Seoul National University School of Dentistry, Seoul, Korea.

ABSTRACT

Objectives: We analyzed gene-expression profiles after 14 day odontogenic induction of human dental pulp cells (DPCs) using a DNA microarray and sought candidate genes possibly associated with mineralization.

Materials and methods: Induced human dental pulp cells were obtained by culturing DPCs in odontogenic induction medium (OM) for 14 day. Cells exposed to normal culture medium were used as controls. Total RNA was extracted from cells and analyzed by microarray analysis and the key results were confirmed selectively by reverse-transcriptase polymerase chain reaction (RT-PCR). We also performed a gene set enrichment analysis (GSEA) of the microarray data.

Results: Six hundred and five genes among the 47,320 probes on the BeadChip differed by a factor of more than two-fold in the induced cells. Of these, 217 genes were upregulated, and 388 were down-regulated. GSEA revealed that in the induced cells, genes implicated in Apoptosis and Signaling by wingless MMTV integration (Wnt) were significantly upregulated.

Conclusions: Genes implicated in Apoptosis and Signaling by Wnt are highly connected to the differentiation of dental pulp cells into odontoblast.

No MeSH data available.


Related in: MedlinePlus