Department of Conservative Dentistry, College of Dentistry, Wonkwang University, Iksan, Korea.
Cryobiology. 2010 Jun;60(3):326-30. doi: 10.1016/j.cryobiol.2010.02.008. Epub 2010 Mar 7.
Gene expression was compared by cDNA microarray analysis in human periodontal ligament (PDL) cells cultured from teeth immediately after extraction and from teeth cryopreserved for 1 week. Twenty healthy collateral premolar teeth without caries and restorations were obtained from 10 young patients, one maxillary and one mandibular premolar from each subject. The teeth from five patients, from two patients, and from three patients out of total 10 patients were used for cDNA microarray assay, for RT-PCR, and for real-time PCR, respectively. One premolar was used immediately after extraction (control), and another premolar was stored in liquid nitrogen at -196 degrees C for 1 week (cryopreserved) from each patient. PDL cells from these teeth were cultured separately through three passages. Total RNA was isolated and gene expression was compared between the cells from control and cryopreserved group out of each subject. The microarray data were validated using the reverse transcription-polymerase chain reaction (RT-PCR) and confirmed by quantitative real-time PCR. The cultured PDL cells from the control and cryopreserved teeth were of similar appearance under an optical microscope. In all subjects the fibroblast growth factor receptor 2 (FGFR2) gene was downregulated in the cells from the cryopreserved tooth. This study shows that cryopreservation of teeth affects the expression of the FGFR2 gene in cultured PDL cells, which is related to cell growth, cell development, and cell-cell signaling.
通过比较人牙周膜（PDL）细胞 cDNA 微阵列分析，比较了立即从提取的牙齿和冷冻保存 1 周的牙齿中培养的基因表达。从 10 名年轻患者中获得了 20 颗健康的侧前磨牙，每个患者各有 1 颗上颌前磨牙和 1 颗下颌前磨牙。总共 10 名患者中的 5 名患者、2 名患者和 3 名患者的牙齿分别用于 cDNA 微阵列分析、RT-PCR 和实时 PCR。每个患者的 1 颗前磨牙立即提取后使用（对照组），另 1 颗前磨牙在 -196°C 液氮中储存 1 周（冷冻组）。从这些牙齿中分别培养 PDL 细胞，通过三个传代进行分离。从对照组和冷冻组的每个患者的牙齿中分离总 RNA，并比较细胞之间的基因表达。使用逆转录聚合酶链反应（RT-PCR）验证微阵列数据，并通过定量实时 PCR 进行确认。在光学显微镜下，从对照和冷冻牙齿培养的 PDL 细胞具有相似的外观。在所有受试者中，冷冻牙齿的成纤维细胞生长因子受体 2（FGFR2）基因在冷冻牙齿来源的细胞中下调。这项研究表明，牙齿的冷冻保存会影响培养的 PDL 细胞中 FGFR2 基因的表达，这与细胞生长、细胞发育和细胞间信号转导有关。
