Induction of Mandibular bone and tooth germ in embryo body formation from mouse and human iPS cells in serum-free cell culture

无血清细胞培养中小鼠和人 iPS 细胞诱导下颌骨和牙胚形成胚胎体

• 批准号: 24659894
• 负责人: OKAMOTO TETSUJI
• 金额: $ 2.41万
• 依托单位: Hiroshima University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Challenging Exploratory Research
• 财政年份: 2012
• 资助国家: 日本
• 起止时间: 2012-04-01 至 2014-03-31
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-24659894/
• 关键词: 無血清培養; エンブリオボディー培養; iPS 細胞; 顎骨誘導; 歯胚誘導; テラトーマ; ３胚葉; 多分化能; 歯髄細胞; ヒトiPS細胞; 無血清培地; フィーダーフリー培養; 胚葉体形成; 口腔顎顔面組織; TGF-β1; マウスiPS細胞; 胚様体培養; 頭部神経系; 歯胚形成; 顎顔面領域; 細胞増殖因子

The miPS cells have been maintained in ESF7 serum-free medium for more than 2 years with an undifferentiated phenotype. An FGF-2 induced miPS cell differentiation into tooth germ progenitor cells. hiPSCs are commonly maintained on feeder cells in medium supplemented with FBS. In this study, we successfully generated hiPSCs from human dental pulp cells using Oct3/4, Sox2, Klf4, c-Myc with retroviral vectors in serum- and feeder-free defined culture conditions. These hiPSC retained the property of selfrenewal, and pluripotency evaluated by differentiation into derivatives of all three primary germ layers by embryo body formation assay and teratoma formation assay. hiPSCs cultured in hESF9 medium absolutely required TGF-beta1 to maintain pluripotency. This simple serum-free adherent monoculture system will allow us to elucidate the cell responses to growth factors under defined conditions and can eliminate the risk might be brought by undefined pathogens.

miPS细胞已在ESF 7无血清培养基中维持超过2年，具有未分化的表型。FGF-2诱导miPS细胞分化为牙胚祖细胞。hiPSC通常维持在补充有FBS的培养基中的饲养细胞上。在这项研究中，我们成功地产生hiPSCs从人牙髓细胞使用Oct 3/4，Sox 2，Klf 4，c-Myc与逆转录病毒载体在血清和饲养层定义的培养条件。这些hiPSC保留了自我更新的性质，并且通过胚胎体形成测定和畸胎瘤形成测定通过分化成所有三个初级胚层的衍生物来评价多能性。在hESF 9培养基中培养的hiPSC绝对需要TGF-β 1来维持多能性。这种简单的无血清贴壁单培养系统将使我们能够阐明细胞在特定条件下对生长因子的反应，并可以消除未知病原体可能带来的风险。

项目成果

Participation of heterodimer formation with integrin αv subunit in the stability of integrin β8 subunit in squamous cell carcinoma cells

整合素αv亚基形成异二聚体参与鳞状细胞癌细胞整合素β8亚基稳定性

• 发表时间: 2013
• 作者: Fujii T.;Hayashido Y.;Hamana T.;Sakaue T. and Okamoto T.
• 通讯作者: Sakaue T. and Okamoto T.

Long-term serial cultivation of mouse induced pluripotent stem cells in serum-free and feeder-free defined medium

• DOI: 10.1387/ijdb.130173to
• 发表时间: 2013-01-01
• 期刊: INTERNATIONAL JOURNAL OF DEVELOPMENTAL BIOLOGY
• 影响因子: 0.7
• 作者: Yamasaki, Sachiko;Nabeshima, Kou;Okamoto, Tetsuji
• 通讯作者: Okamoto, Tetsuji

Biomedical advances from tissue culture

• DOI: 10.1007/s10616-013-9591-1
• 发表时间: 2013-12-01
• 期刊: CYTOTECHNOLOGY
• 影响因子: 2.2
• 作者: Okamoto，Tetsuji;Sato，J. Denry;Sato，Gordon H.
• 通讯作者: Sato，Gordon H.

Generation of disease-specific human induced pluripotent stem (iPS) cells from dental pulp cells of a patient with Cleidocranial dysplasia in serum-, and feeder-free culture

在无血清和无饲养层培养中，从锁骨颅骨发育不良患者的牙髓细胞中生成疾病特异性人诱导多能干 (iPS) 细胞

• 发表时间: 2016
• 作者: A. Hamada;S. Yamasaki;E. Akagi;H. Nakatao;F. Obayashi;T. Fukutani;K. Koizumi;T. Hamana;T. Okamoto
• 通讯作者: T. Okamoto

Generation of induced pluripotent stem cells from dental pulp cells in serum-free and feeder-free culture condition

无血清、无饲养层培养条件下牙髓细胞诱导多能干细胞的生成

• 发表时间: 2013
• 作者: Taguchi Y.;Yamasaki S.;ShimamotoA.;Tahara H. and Okamoto T
• 通讯作者: Tahara H. and Okamoto T