Platform for induction and maintenance of transgene-free hiPSCs resembling ground state pluripotent stem cells.
Platform for induction and maintenance of transgene-free hiPSCs resembling ground state pluripotent stem cells.
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DOI：
10.1016/j.stemcr.2014.01.014
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发表时间：
2014-03-11
影响因子：
5.9
通讯作者：
Flynn, Peter
中科院分区：
文献类型：
--
作者：
Valamehr, Bahram;Robinson, Megan;Abujarour, Ramzey;Rezner, Betsy;Vranceanu, Florin;Le, Thuy;Medcalf, Amanda;Lee, Tom Tong;Fitch, Michael;Robbins, David;Flynn, Peter
细胞建库、疾病建模和细胞治疗应用对hiPSC技术提出了越来越高的要求。具体而言，无足迹hiPSC的高通量衍生及其在允许规模化生产的系统中的扩增仍然具有技术挑战性。在这里，我们描述了一种使用阶段特异性培养基组合物中的小分子途径抑制剂快速、平行产生、选择和扩增hiPSC的平台。该平台支持在完全无饲养层的环境中仅使用OCT 4/SOX 2/SV 40 LT组合（0.5%-4.0%，在第12天和第16天之间）进行有效和加速的游离型重编程。所得hiPSC不含转基因，易于培养，并作为单细胞扩增，同时保持同质和基因组稳定的多能群体。在所述培养基组合物中产生或维持的hiPSC表现出与多能性基态相关的性质。该系统的简单性和稳健性允许高通量生成和快速扩展适用于工业和临床级用途的均匀hiPSC产品。FRM支持高效的最小因子附加型重编程通过高分辨率单细胞分选实现hiPSC的克隆衍生FMM支持长期维持基因组稳定、无转基因的hiPSC维持类似基态的同质多能培养细胞治疗和建库应用对hiPSC技术提出了越来越高的要求。在这里，Flynn及其同事展示了一种简单的多重选择和维持平台，该平台利用阶段特异性小分子培养基添加剂和流式细胞术分选来衍生基因组稳定且类似于基态多能干细胞的最小因子附加型hiPSC。该平台可以作为产生临床相关hiPSC的途径。
Cell banking, disease modeling, and cell therapy applications have placed increasing demands on hiPSC technology. Specifically, the high-throughput derivation of footprint-free hiPSCs and their expansion in systems that allow scaled production remains technically challenging. Here, we describe a platform for the rapid, parallel generation, selection, and expansion of hiPSCs using small molecule pathway inhibitors in stage-specific media compositions. The platform supported efficient and expedited episomal reprogramming using just OCT4/SOX2/SV40LT combination (0.5%–4.0%, between days 12 and 16) in a completely feeder-free environment. The resulting hiPSCs are transgene-free, readily cultured, and expanded as single cells while maintaining a homogeneous and genomically stable pluripotent population. hiPSCs generated or maintained in the media compositions described exhibit properties associated with the ground state of pluripotency. The simplicity and robustness of the system allow for the high-throughput generation and rapid expansion of a uniform hiPSC product that is applicable to industrial and clinical-grade use. FRM supports highly efficient minimal factor episomal reprogramming Clonal derivation of hiPSCs through high-resolution single-cell sorting FMM supports long-term maintenance of genomically stable, transgene-free hiPSCs Maintenance of homogeneous pluripotent culture resembling the ground state Cell therapy and banking applications have placed increasing demands on hiPSC technology. Here, Flynn and colleagues demonstrate a simple multiplex selection and maintenance platform utilizing stage-specific small molecule media additives and flow cytometry sorting to derive minimal factor episomal hiPSCs that are genomically stable and resemble ground state pluripotent stem cells. This platform may serve as a path to generate clinically relevant hiPSCs.
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影响因子：
48
作者：
Lin, Tongxiang;Ambasudhan, Rajesh;Yuan, Xu;Li, Wenlin;Hilcove, Simon;Abujarour, Ramzey;Lin, Xiangyi;Hahm, Heung Sik;Hao, Ergeng;Hayek, Alberto;Ding, Sheng
通讯作者：
Ding, Sheng
影响因子：
64.5
作者：
Hanna JH;Saha K;Jaenisch R
通讯作者：
Jaenisch R
影响因子：
23.9
作者：
Saha K;Jaenisch R
通讯作者：
Jaenisch R
间充质到上皮的转变启动并且是小鼠成纤维细胞核重编程所必需的
影响因子：
23.9
作者：
Li, Ronghui;Liang, Jialiang;Pei, Duanqing
通讯作者：
Pei, Duanqing
影响因子：
4.6
作者：
Abujarour, Ramzey;Valamehr, Bahram;Robinson, Megan;Rezner, Betsy;Vranceanu, Florin;Flynn, Peter
通讯作者：
Flynn, Peter