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Proliferation of dental follicle-derived cell populations in heat-stress conditions.

ABSTRACT: Isolation and purification of adult stem cells (ASC) are a great challenge. Our objectives were to determine whether ASC are more heat-tolerant than non-stem cells, and to explore if ASC could be enriched by heat-stress treatments.Rat dental follicle cells were cultured in a variety of media to obtain either a heterogeneous cell population (H-DFC) consisting of stem cells and non-stem cells, or a homogenous cell population (DFC) containing non-stem cells only. Real-time RT-PCR was conducted to compare expression of heat-shock proteins (HSPs) between the two populations. To study heat tolerance, H-DFC and DFC were incubated under heat-stress conditions and cell proliferation was evaluated by alamar blue reduction assay. Furthermore, cells resulting from heat-stress treatments were evaluated for differentiation capability and expression of stem cell markers.H-DFC expressed higher levels of HSP110, HSP70s and HSP27s than did DFC. H-DFC increased levels of proliferation at 40 °C compared to controls grown at 37 °C; no significant reduction in proliferation occurred at temperatures below 40.5 °C. In contrast, DFC showed significant reduction in proliferation under all heat-stress treatments. Heat-stressed H-DFC had increased differentiation capability and increased expression of stem cell markers.Stem cells appear to be more tolerant to heat stress than non-stem cells. Incubation of a heterogeneous cell population in heat-stress conditions resulted in increased stem cell numbers.

SUBMITTER: Yao S

PROVIDER: S-EPMC3187570 | biostudies-literature | 2011 Oct

REPOSITORIES: biostudies-literature

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Proliferation of dental follicle-derived cell populations in heat-stress conditions.

Yao S S Gutierrez D L DL He H H Dai Y Y Liu D D Wise G E GE

Cell proliferation 20111001 5

<h4>Objectives</h4>Isolation and purification of adult stem cells (ASC) are a great challenge. Our objectives were to determine whether ASC are more heat-tolerant than non-stem cells, and to explore if ASC could be enriched by heat-stress treatments.<h4>Materials and methods</h4>Rat dental follicle cells were cultured in a variety of media to obtain either a heterogeneous cell population (H-DFC) consisting of stem cells and non-stem cells, or a homogenous cell population (DFC) containing non-ste ...[more]

PMID: 21951291

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Project description:Bone formation is important for the reconstruction of bone-related structures in areas that have been damaged by inflammation. Inflammatory conditions such as those that occur in patients with rheumatoid arthritis, cystic fibrosis, and periodontitis have been shown to inhibit osteoblastic differentiation. This study focussed on dental follicle stem cells (DFSCs), which are found in developing tooth germ and participate in the reconstruction of alveolar bone and periodontal tissue in periodontal disease. After bacterial infection of inflamed dental tissue, the destruction of bone was observed. Currently, little is known about the relationship between the inflammatory environment and bone formation. Osteogenic differentiation of inflamed DFSCs resulted in decreased alkaline phosphatase (ALP) activity and alizarin red S staining compared to normal DFSCs. Additionally, in vivo transplantation of inflamed and normal DFSCs demonstrated severe impairment of osteogenesis by inflamed DFSCs. Protein profile analysis via liquid chromatography coupled with tandem mass spectrometry was performed to analyse the differences in protein expression in inflamed and normal tissue. Comparison of inflamed and normal DFSCs showed significant changes in the level of expression of transforming growth factor (TGF)-β2. Porphyromonas gingivalis (P.g.)-derived lipopolysaccharide (LPS) was used to create in vitro inflammatory conditions similar to periodontitis. The osteogenic differentiation of LPS-treated DFSCs was suppressed, and the cells displayed low levels of TGF-β1 and high levels of TGF-β2. DFSCs treated with TGF-β2 inhibitors showed significant increases in alizarin red S staining and ALP activity. TGF-β1 expression was also increased after inhibition of TGF-β2. By examining inflamed DFSCs and LPS-triggered DFSCs, these studies showed both clinically and experimentally that the increase in TGF-β2 levels that occurs under inflammatory conditions inhibits bone formation.

Dataset Information

Proliferation of dental follicle-derived cell populations in heat-stress conditions.

Publications

Proliferation of dental follicle-derived cell populations in heat-stress conditions.

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