Establishment of safe and efficient differentiation system using automatic removal vector

利用自动去除向量建立安全高效的分化系统

• 批准号: 24700474
• 负责人: NISHIMURA Ken
• 金额: $ 2.83万
• 依托单位: University of Tsukuba
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Young Scientists (B)
• 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/grant/KAKENHI-PROJECT-24700474/
• 关键词: 分化; 多能性幹細胞; 残存未分化細胞; SeVdpベクター; miRNA; 持続発現型RNAベクター; 安全性

When we induce differentiated cells from pluripotent stem cells, the risks of tumor formation by remained undifferentiated cells must be concerned. To reduce such a risk, we tried to establish a system for elimination of undifferentiated cells in the differentiated tissue using our SeVdp vector system. The SeVdp vector which has the target sequence of neuron-specific miRNA was infected to mouse iPS cells, and then they were differentiated to neuron. As a result, the vector were automatically removed from neuron while it was persistently infected in undifferentiated cells. But when we tried to select differentiated cells using negative selection gene from the vector, not only undifferentiated cells but also differentiated cells were killed by the treatment. We plan to investigate other method of differentiation and negative selection to selectively eliminate the undifferentiated cells.

当我们从多能干细胞中诱导分化细胞时，必须考虑未分化细胞形成肿瘤的风险。为了降低这种风险，我们尝试使用我们的SeVdp载体系统建立用于消除分化组织中未分化细胞的系统。将含有神经元特异性miRNA靶序列的SeVdp载体感染小鼠iPS细胞，诱导其向神经元分化。结果，该载体在未分化细胞中持续感染时，自动从神经元中去除。但是当我们尝试使用来自载体的负选择基因来选择分化的细胞时，不仅未分化的细胞而且分化的细胞都被处理杀死。我们计划研究其他的分化和负选择方法来选择性地去除未分化的细胞。

项目成果

初期化が途中で停止した細胞を用いたiPS細胞誘導過程の解析

使用重编程中途停止的细胞分析 iPS 细胞诱导过程

• 发表时间: 2014
• 作者: Higa M.;Chih-Chiang Chang;Roche C;Struk A. M.;Farmer K. W.;Wright T. W.;Banks S. A.;西村健
• 通讯作者: 西村健

QUALITY CONTROL OF IPS CELLS BY UNIQUE GENE TRANSFER SYSTEM

通过独特的基因转移系统对 IPS 细胞进行质量控制

• 发表时间: 2012
• 作者: 井上祐貴;坂田翔;石原一彦;K. Nishimura
• 通讯作者: K. Nishimura

Taking snap shots during iPSC production by SeVdp vectors

在 iPSC 生产过程中使用 SeVdp 载体拍摄快照

• 发表时间: 2013
• 作者: Higa M.;Chih-Chiang Chang;Roche C;Struk A.M.;Farmer K.W.;Wright T.W.;Banks S.A.;K. Nishimura
• 通讯作者: K. Nishimura

SeVdpベクターを用いたiPS細胞の品質調節メカニズムの解析

利用SeVdp载体分析iPS细胞的质量调控机制

• 发表时间: 2013
• 作者: M. Higa;H. Tanino;Y. Yamagami;M. Abo;and S. Kakunai;西村健
• 通讯作者: 西村健

Klf4遺伝子発現量調節による初期化が途中で停止した細胞の誘導

诱导细胞在调节 Klf4 基因表达水平的过程中停止重编程

• 发表时间: 2013
• 作者: Masaru Higa;Fusako Kawabata;Masayoshi Abo;and Satoshi Kakunai,;西村健
• 通讯作者: 西村健