Zhang Chao, Cheng Xiang-rong
School of Stomatology, Wuhan University, Wuhan, China.
Zhonghua Kou Qiang Yi Xue Za Zhi. 2013 Feb;48(2):96-101.
To isolate and characterize the dental follicle cells (DFC) from dental follicle (DF) tissues of normal human impacted third molars.
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).
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.
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.
从正常人类阻生第三磨牙的牙囊组织中分离并鉴定牙囊细胞（DFC）。
从健康年轻人类阻生第三磨牙的牙囊组织中分离DFC。采用有限稀释培养法评估DFC的集落形成效率。通过免疫组织化学分析检测DFC中Stro-1、Notch-1和巢蛋白的表达。对原代DFC培养物进行多种处理模式：成骨、成脂和软骨分化。通过甲基噻唑基四氮唑（MTT）法比较DFC和牙周膜细胞（PDLC）的增殖能力。通过逆转录聚合酶链反应（RT-PCR）评估DFC和PDLC中腱生蛋白-N和F-腱蛋白的表达。
大多数DFC为纺锤形成纤维细胞样细胞。DFC培养物从第1代细胞形成集落，集落形成效率（CFE）频率为3.70%。部分DFC对Stro-1呈阳性染色，几乎所有DFC对Notch-1和巢蛋白呈阳性染色。DFC培养物在进行21天的命运特异性诱导后表现出多能特性。成骨诱导后检测到茜素红阳性凝聚结节，成脂诱导后产生油红阳性脂滴，软骨诱导后通过免疫组织化学显示Ⅱ型胶原。在第3天和第5天，DFC（0.20±0.01，0.51±0.09）比PDLC（0.16±0.03，0.47±0.07）表现出更好的细胞活性（P>0.05）。在第7天，DFC（1.03±0.11）的增殖率高于PDLC（0.93±0.09）（P<0.05）。RT-PCR结果显示腱生蛋白-N在DFC中不表达，但在PDLC中中度表达。F-腱蛋白在DFC中强烈表达，而在PDLC中不表达。
来自外胚间充质组织的DFC具有良好的活力，并且含有与间充质干细胞相似的细胞。它可能用作牙周组织再生的新型细胞来源。
