Basic Fibroblast Growth Factor Regulates REX1 Expression Via IL‐6 In Stem Cells Isolated From Human Exfoliated Deciduous Teeth

Abstract

Basic fibroblast growth factor (bFGF) regulates pluripotent marker expression and cellular differentiation in various cell types. However, the mechanism by which bFGF regulates REX1 expression in stem cells, isolated from human exfoliated deciduous teeth (SHEDs) remains unclear. The aim of the present study was to investigate the regulation of REX1 expression by bFGF in SHEDs. SHEDs were isolated and characterized. Their mRNA and protein expression levels were determined using real-time polymerase chain reaction and enzyme-linked immunosorbent assay, respectively. In some experiments, chemical inhibitors were added to the culture medium to impede specific signaling pathways. Cells isolated from human exfoliated deciduous tooth dental pulp tissue expressed mesenchymal stem cell surface markers (CD44, CD73, CD90, and CD105). These cells differentiated into osteogenic and adipogenic lineages, when appropriately induced. Treating SHEDs with bFGF induced REX1 mRNA expression and this effect was attenuated by pretreatment with FGFR or Akt inhibitors. Cycloheximide pretreatment also inhibited the bFGF-induced REX1 expression, implying the involvement of intermediate molecule(s). Further, the addition of an IL-6 neutralizing antibody attenuated the bFGF-induced REX1 expression by SHEDs. In conclusion, bFGF enhanced REX1 expression by SHEDs via the FGFR and Akt signaling pathways. Moreover, IL-6 participated in the bFGF-induced REX1 expression in SHEDs. J. Cell. Biochem. 118: 1480-1488, 2017. © 2016 Wiley Periodicals, Inc.

Dental properties, ultrastructure, and pulp cells associated with a novel DSPP mutation

“…Cells were isolated from the remaining dental pulp tissues of an extracted deciduous incisor from the proband and unaffected controls. Tissue explant technique was employed for cell isolation according to previous publications (Nowwarote, Pavasant, & Osathanon, ; Nowwarote, Sukarawan, Pavasant, & Osathanon, ). Cells were cultured in Dulbecco's Modified Eagle Medium (DMEM, Gibco ® ) containing 10% foetal bovine serum (FBS, Gibco ® , USA), 100 unit/ml penicillin, 100 μg/ml streptomycin, 5 μg/ml amphotericin B, 2 mM l‐glutamine (Glutamax ® ) at 37°C in 5%CO 2 .…”

“…Cells were maintained in growth medium supplemented with 50 μg/ml ascorbic acid, 10 mM beta‐glycerophosphate, and 100 nM dexamethasone. Mineral deposition was examined by alizarin red S staining according to previous publications (Nowwarote et al., , ). The expression of osteogenic marker genes was evaluated using quantitative polymerase chain reaction.…”

Stem Cells from Human Exfoliated Deciduous Teeth: Biology and Therapeutic Potential

“…Several studies have reported that SHEDs can be induced into adipogenic lineage [6,[32][33][34]. After cultured in an adipogenic medium, SHEDs' morphology changed from spindle-like to polygonal shapes and lipid vacuoles were observed, along with the increased in PPARγ2 and LPL mRNA [32].…”

“…Further, bFGF promotes colony-forming unit ability in SHEDs isolated from inflamed dental pulp tissues [42]. For the regulatory mechanism, it has been demonstrated that bFGF regulated REX-1 expression in SHEDs via FGFR and Akt pathway [34]. IL-6 is also shown to involve in bFGF induced REX-1 expression as pre-treatment with antibody against IL-6 attenuates REX-1 expression [34].…”

Biphasic Effect of ATP on In Vitro Mineralization of Dental Pulp Cells

“…The expression of the surface markers CD44, CD90, CD105, and CD45 were evaluated. The multilineage differentiation protocols were performed according to previous reports [Govitvattana et al, 2013;Nowwarote et al, 2015Nowwarote et al, , 2017. Briefly, cells were cultured in OM (GM supplemented with 50 mg/ml ascorbic acid (Sigma-Aldrich Chemical), 100 nM dexamethasone (Sigma-Aldrich Chemical), and 10 mM b-glycerophosphate (BGP) (Sigma-Aldrich Chemical)) or adipogenic induction medium (AM) (GM supplemented with 0.1 mg/ml insulin, 1 mM dexamethasone, 1 mM IBMX, and 0.2 mM indomethacin).…”