Established efficiency of iPS cells derived from impacted supernumerary tooth considering bio-recycle

考虑生物回收，确定源自受影响多生牙的 iPS 细胞的效率

• 批准号: 23792364
• 负责人: SAITO Yoko
• 金额: $ 2.75万
• 依托单位: Niigata University (2012)Kagoshima University (2011)
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Young Scientists (B)
• 财政年份: 2011
• 资助国家: 日本
• 起止时间: 2011 至 2012
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-23792364/
• 关键词: 小児歯科学; 再生医学; バイオリサイクル; 歯; iPS細胞; iＰＳ細胞

Feeder cells are generally required for establishing and maintaining embryonic stem (ES) cells/inducible pluripotent stem (iPS) cells. Mouse embryonic fibroblasts (MEFs) isolated from fetuses and STO mouse stromal cell line are the most widely used cells for this purpose. The aim of this study was to determine which cells are suitable for establishing iPS cells from human deciduous tooth dental pulp cells (HDDPCs) and supernumerary teeth (SNT). Primarily cultured HDDPCs and SNT (5 × 104) wereco-transfected with 3 plasmids containing human OCT3/4, SOX2/KLF4, or LMYC/LIN28 together with pmaxGFP (used as a reporter) by using the NeonR Transfection system, a novel electroporation method involving a capillary and wire-type electrode. They were then cultured in a medium specified for maintaining ES cells for 15 days. Colonies were collected by trypsinization and re-seeded onto mitomycin C-treated MEFs or STO cells. The colonies were serially passaged for up to 26 passages. During this period, colony morphology was assessed to determine whether they exhibit ES-like morphology and alkaline phosphatase activity to evaluate the state of HDDPC reprogramming. HDDPCs maintained on MEFs were transformed to iPS cells with several undifferentiated properties, but those on STO cells failed. Established HDDPC-derived iPS cells grown on MEFs were successfully maintained on STO cells without loss of their pluripotent nature. Our results indicate that HDDPCs can be reprogrammed using reprogramming factor, and that MEFs are better than STO cells as feeder cells for establishing and maintaining HDDPC-derived iPS cells, suggesting that the choice of feeders is a key factors enabling efficient generation of iPS cells.

饲养层细胞通常是建立和维持胚胎干细胞/可诱导多能干细胞所必需的。从胚胎中分离的小鼠胚胎成纤维细胞(MEF)和STO小鼠基质细胞系是应用最广泛的细胞。本研究的目的是确定哪些细胞适合从人乳牙牙髓细胞(HDDPC)和多生牙(SNT)建立iPS细胞。将含有人Oct3/4、SOX2/KLF4和LMYC/LIN28的3种真核表达载体与pMaxGFP(以pMaxGFP为报告载体)通过NeonR电穿孔系统(一种新型的毛细管和线状电极)共同导入原代培养的HDDPC和SNT(5×104)。然后，他们在一种特定的维持ES细胞的培养液中培养15天。通过胰酶消化收集集落，重新接种到丝裂霉素C处理的MEF或STO细胞上。蜂群经过长达26次的连续传代。在此期间，评估集落形态以确定它们是否表现出ES样形态和碱性磷酸酶活性，以评估HDDPC重编程的状态。保持在MEF上的HDDPC被转化为具有多种未分化特性的iPS细胞，而在STO细胞上的HDDPC则失败了。在MEF上生长的已建立的HDDPC来源的iPS细胞成功地维持在STO细胞上，而不会失去其多能性。我们的结果表明，可以使用重编程因子对HDDPC进行重编程，并且MEF作为饲养层细胞比STO细胞更适合于建立和维持HDDPC来源的iPS细胞，提示饲养层的选择是高效生成iPS细胞的关键因素。

项目成果

小児における経鼻挿管チューブの固定長に関する研究

儿童鼻插管固定长度的研究

• 发表时间: 2011
• 作者: 真鍋庸三;齊藤陽子;遠矢明菜;下松孝太;糀谷淳;椙山加綱
• 通讯作者: 椙山加綱

Choice of feeders is important for the preparation of iPS cells from primarily cultured human deciduous tooth dental pulp cells

饲养层的选择对于从原代培养的人乳牙牙髓细胞制备 iPS 细胞很重要

• 发表时间: 2013
• 作者: Iwase Y;Saitoh I;Okamoto A;Nakakura-Ohshima K;Inada E;Yamada C;Takemoto Y;Yamasaki Y;Hayasaki H;村上智哉,齊藤一誠,稲田絵美,岩瀬陽子,長谷川大子,窪田直子,松本祐子,大島邦子,岡暁子,山崎要一,早崎治明
• 通讯作者: 村上智哉,齊藤一誠,稲田絵美,岩瀬陽子,長谷川大子,窪田直子,松本祐子,大島邦子,岡暁子,山崎要一,早崎治明

口腔外科長時間手術において、グルコース・インスリン投与による血糖値管理が術後白血球数・C反応性タンパク値・体温および退院日数に及ぼす影響

长期口腔外科手术期间通过葡萄糖和胰岛素管理血糖水平对术后白细胞计数、C反应蛋白水平、体温和出院时间的影响

• 发表时间: 2011
• 作者: 遠矢明菜;糀谷淳;真鍋庸三;齊藤陽子;椙山加綱
• 通讯作者: 椙山加綱

加速度脈波解析におけるc/aは何を意味するのか?

加速脉搏波分析中的 c/a 是什么意思？

• 发表时间: 2011
• 作者: 齊藤陽子;糀谷淳;真鍋庸三;遠矢明菜;椙山加綱
• 通讯作者: 椙山加綱

当大学病院小児歯科における静脈内鎮静法を用いた歯科治療の経時的臨床統計

我们大学医院儿科牙科使用静脉镇静牙科治疗随时间的临床统计

• 发表时间: 2011
• 作者: 窪田直子;深水篤;齊藤一誠;岩崎智憲;長谷川大子;伊藤千晶;村上大輔;糀谷淳;齊藤陽子;遠矢明菜;椙山加綱;山崎要一
• 通讯作者: 山崎要一