The modulatory mechanism of bone morphogenetic protein 4 on the expression of reprogramming markers in human dental pulp cells

doi:10.3877/cma.j.issn.1674-1366.2011.05.001

Abstract

Abstract:

Objective

The purpose of this study was to investigate the modulatory role of exogenous bone morphogenetic protein 4 （BMP-4） on the growth and the expression of reprogramming markers in human dental pulp cells with long term in vitro culture.

Methods

Cell counting was applied to the DPCs growing our from human dental pulp tissue with 4 w in vitro culture， either stimulated with recombinant human BMP-4（rhBMP-4） or without. Immunofluorescent staining and quantitative Realtime PCR were applied to investigate the expression of Oct-4， Sox-2 and c-Myc expression on DPCs at passage 2 and 7 of rhBMP-4 induced and control group.

Results

The number of out growing DPCs after 4 w in vitro cultured with rhBMP-4 was statistically higher than the one from control group （P＜0.05）. Oct-4， Sox-2 and c-Myc showed a similar expressing pattern in DPCs from the rhBMP-4 induced and control group at passage 2， which showed their positive nucleus expression. Whereas they lost nucleus location and showed weak cytoplasm expression in DPCs at passage 7 in control group， albeit remained necleus location in DPCs at passage 7 from the rhBMP-4 induced group. The mRNA expression of Oct-4， Sox-2 and c-Myc was similar in DPCs from both rhBMP-4 induced and control group （P＞0.05）， whereas their mRNA expression were all sinificantly higher in DPCs from rhBMP-4 induced group than the control group（P＜0.05）.

Conclusions

BMP-4 can stimulate the growth of DPCs， and enhance the expression of Oct-4， Sox-2 and c-Myc in DPCs with long term in vitro culture， implying BMP-4 may regulate the undifferentiated capacity of DPCs with in vitro culture.

Key words: Bone morphology protein-4, Dental pulp cells, Pluripotency genes

Lu LIU, Xi WEI, Jun-qi LING, Fang ZHANG, Li-ping WU. The modulatory mechanism of bone morphogenetic protein 4 on the expression of reprogramming markers in human dental pulp cells[J]. Chinese Journal of Stomatological Research(Electronic Edition), 2011, 5(05): 455-462.

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Figures/Tables 6

References 19

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基因	引物序列
Oct-4	Forward:5′-GCT CGA GAA GGA TGT GGT C-3′
	Reverse:5′-ATC CTC TCG TTG TGC ATA GTC G-3′
Sox-2	Forward:5′-CAC TGF CCC TCT CAC ACA TG-3′
	Reverse:5′-CCC ATT TCC CTC GTT TTT CTT-3′
c-Myc	Forward:5′-CGT CTC CAC ACA TCA GCA CAA-3′
	Reverse:5′-TCT TGG CAG CAG GAT AGT CCT T-3′
18s	Forward:5′-TCG GAA CTG AGG CCA TGA TTA AG-3′
	Reverse:5′-TCT TCG AAC CTC CGA CTT TCG-3′

基因	引物序列
Oct-4	Forward:5′-GCT CGA GAA GGA TGT GGT C-3′
	Reverse:5′-ATC CTC TCG TTG TGC ATA GTC G-3′
Sox-2	Forward:5′-CAC TGF CCC TCT CAC ACA TG-3′
	Reverse:5′-CCC ATT TCC CTC GTT TTT CTT-3′
c-Myc	Forward:5′-CGT CTC CAC ACA TCA GCA CAA-3′
	Reverse:5′-TCT TGG CAG CAG GAT AGT CCT T-3′
18s	Forward:5′-TCG GAA CTG AGG CCA TGA TTA AG-3′
	Reverse:5′-TCT TCG AAC CTC CGA CTT TCG-3′

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