Isolation and characterization of dental follicle cells from adult human dental follicle tissues / 中华口腔医学杂志

ABSTRACT

<p><b>OBJECTIVE</b>To isolate and characterize the dental follicle cells (DFC) from dental follicle (DF) tissues of normal human impacted third molars.</p><p><b>METHODS</b>DFC were isolated from the DF tissues of healthy young human impacted third molars. A limited dilution culture was used to assess DFC colony-forming efficiency. The expressions of Stro-1, Notch-1 and nestin in DFC were detected by immunohistochemistry analysis. The primary DFC cultures were subjected to a variety of treatment modes osteogenic, adipogenic and chondrogenic differentiation. DFC and periodontal ligament cells (PDLC) proliferation abilities were compared by methyl thiazolyl tetrazolium (MTT) assay. The expressions of tenascin-N and F-spondin in DFC and PDLC were evaluated by reverse transcriptase polymerase chain reaction (RT-PCR).</p><p><b>RESULTS</b>Most DFC were spindle fibroblast-like cells. DFC cultures formed colonies from passage 1 cells and the frequency of colony forming efficiency (CFE) was 3.70%. Some of the DFC were stained positively for Stro-1 and almost all the DFC were stained positively for Notch-1 and nestin. DFC cultures displayed multipotential characteristics following fate-specific inductions for 21 days. Alizarin red positive condensed nodules were detected following osteogenic induction, oil red-positive lipid vacuoles were generated using adipogenic induction and collagen-II was revealed following chondrogenic induction by immunohistochemistry. On day 3 and 5, DFC (0.20 ± 0.01, 0.51 ± 0.09) showed a better cell activity than PDLC (0.16 ± 0.03, 0.47 ± 0.07) (P > 0.05). On day 7, DFC (1.03 ± 0.11) exhibited a higher proliferation rate than PDLC (0.93 ± 0.09) (P < 0.05). RT-PCR results showed that tenascin-N was not expressed in DFC, but expressed moderately in PDLC. F-spondin was expressed strongly in DFC, while not expressed in PDLC.</p><p><b>CONCLUSIONS</b>DFC from ectomesenchymal tissues showed a good viability and contained cells similar to the mesenchymal stem cells. It may be used as a novel cell source for periodontium regeneration.</p>