A cost-effective and efficient reprogramming platform for large-scale production of integration-free human induced pluripotent stem cells in chemically defined culture.
A cost-effective and efficient reprogramming platform for large-scale production of integration-free human induced pluripotent stem cells in chemically defined culture.
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DOI：
10.1038/srep11319
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发表时间：
2015-06-11
影响因子：
4.6
通讯作者：
Chen G
中科院分区：
文献类型：
--
作者：
Beers J;Linask KL;Chen JA;Siniscalchi LI;Lin Y;Zheng W;Rao M;Chen G
限制采用IPSC技术的因素包括开发生产线的成本以及表征和存储这些生产线所需的时间周期，特别是在使用无集成、无馈线和无Xeno元件的情况下。在这份手稿中，我们描述了我们的优化程序，使一名技术人员能够以可靠和可重复的方式，以24-96井的格式一次制作20-40条线。改进贯穿整个工作流程，包括使用RNA病毒，降低试剂的细胞毒性，开发改进的转染率和冷冻效率，修改手动菌落挑选步骤，提高传代效率，制定早期成功标准。这些改装使我们每年可以制作200多条具有良好特点的生产线。
Factors limiting the adoption of iPSC technology include the cost of developing lines and the time period that it takes to characterize and bank them, particularly when integration free, feeder free, and Xeno-free components are used. In this manuscript we describe our optimization procedure that enables a single technician to make 20–40 lines at a time in a 24–96 well format in a reliable and reproducible fashion. Improvements spanned the entire workflow and included using RNA virus, reducing cytotoxicity of reagents, developing improved transfection and freezing efficiencies, modifying the manual colony picking steps, enhancing passaging efficiency and developing early criteria of success. These modifications allowed us to make more than two hundred well-characterized lines per year.
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影响因子：
14.8
作者：
通讯作者：
--
影响因子：
12.3
作者：
Gentleman RC;Carey VJ;Bates DM;Bolstad B;Dettling M;Dudoit S;Ellis B;Gautier L;Ge Y;Gentry J;Hornik K;Hothorn T;Huber W;Iacus S;Irizarry R;Leisch F;Li C;Maechler M;Rossini AJ;Sawitzki G;Smith C;Smyth G;Tierney L;Yang JY;Zhang J
通讯作者：
Zhang J
影响因子：
48
作者：
Burridge, Paul W.;Matsa, Elena;Shukla, Praveen;Lin, Ziliang C.;Churko, Jared M.;Ebert, Antje D.;Lan, Feng;Diecke, Sebastian;Huber, Bruno;Mordwinkin, Nicholas M.;Plews, Jordan R.;Abilez, Oscar J.;Cui, Bianxiao;Gold, Joseph D.;Wu, Joseph C.
通讯作者：
Wu, Joseph C.
DOI：
10.1126/science.1172482
发表时间：
2009-05-08
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Yu J;Hu K;Smuga-Otto K;Tian S;Stewart R;Slukvin II;Thomson JA
通讯作者：
Thomson JA
影响因子：
23.9
作者：
Warren, Luigi;Manos, Philip D.;Ahfeldt, Tim;Loh, Yuin-Han;Li, Hu;Lau, Frank;Ebina, Wataru;Mandal, Pankaj K.;Smith, Zachary D.;Meissner, Alexander;Daley, George Q.;Brack, Andrew S.;Collins, James J.;Cowan, Chad;Schlaeger, Thorsten M.;Rossi, Derrick J.
通讯作者：
Rossi, Derrick J.