Isolation and characterization of dental follical cells from adult human dental follicle tissues
10.3760/cma.j.issn.1002-0098.2013.02.008
- VernacularTitle:成体人牙囊细胞的分离培养及生物学特性研究
- Author:
Chao ZHANG
1
;
Xiang-Rong CHENG
Author Information
1. 首都医科大学口腔医学院修复科
- Keywords:
Dental sac;
Stem cells;
Cell culture techniques;
Tissue engineering
- From:
Chinese Journal of Stomatology
2013;48(2):96-101
- CountryChina
- Language:Chinese
-
Abstract:
Objective To isolate and characterize the dental follical cells(DFC) from dental follicle (DF) tissues of normal human impacted third molars.Methods 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 thiazoloyl 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).Results 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 re vealed 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.Conclusions 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.
