A systematic evaluation of integration free reprogramming methods for deriving clinically relevant patient specific induced pluripotent stem (iPS) cells.
A systematic evaluation of integration free reprogramming methods for deriving clinically relevant patient specific induced pluripotent stem (iPS) cells.
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DOI：
10.1371/journal.pone.0081622
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Nathwani AC
中科院分区：
文献类型：
--
作者：
Goh PA;Caxaria S;Casper C;Rosales C;Warner TT;Coffey PJ;Nathwani AC
使用相同的一组亲本细胞对用于产生诱导多能干（iPS）细胞的三种不同方法进行系统评价，以开发一种用于体细胞重编程的不依赖饲养层和无异种的方法，该方法可以转移到GMP环境中。当使用BJ成纤维细胞系时，用基于mRNA的方法观察到最高的重编程效率（起始细胞的1.89%），其几乎是用逆转录病毒（0.2%）和附加型质粒（0.10%）方法观察到的重编程效率的20倍。标准表征测试未揭示使用各种方法衍生的iPS系之间的多能性标志物阵列的任何差异。然而，当使用相同的方法对三种不同的原代成纤维细胞系进行重编程时，两种来源于患有快速发作的帕金森氏症肌张力障碍的患者，一种来源于老年健康志愿者，我们始终观察到附加型质粒方法的重编程效率更高，与逆转录病毒方法相比，其高4倍，比mRNA方法高50倍以上。此外，对于质粒重编程方案，重组玻连蛋白和synthemax®可以与市售的、完全限定的、无异种的必需8培养基一起使用，而不会显著影响重编程效率。为了证明该方案的稳健性，我们重新编程了另外2个原代患者细胞系，一个患有视网膜色素变性，另一个患有帕金森病。我们相信，我们已经优化了一种简单且可重复的方法，可用作开发GMP方案的起点，这是产生临床相关患者特异性iPS细胞的先决条件。
A systematic evaluation of three different methods for generating induced pluripotent stem (iPS) cells was performed using the same set of parental cells in our quest to develop a feeder independent and xeno-free method for somatic cell reprogramming that could be transferred into a GMP environment. When using the BJ fibroblast cell line, the highest reprogramming efficiency (1.89% of starting cells) was observed with the mRNA based method which was almost 20 fold higher than that observed with the retrovirus (0.2%) and episomal plasmid (0.10%) methods. Standard characterisation tests did not reveal any differences in an array of pluripotency markers between the iPS lines derived using the various methods. However, when the same methods were used to reprogram three different primary fibroblasts lines, two derived from patients with rapid onset parkinsonism dystonia and one from an elderly healthy volunteer, we consistently observed higher reprogramming efficiencies with the episomal plasmid method, which was 4 fold higher when compared to the retroviral method and over 50 fold higher than the mRNA method. Additionally, with the plasmid reprogramming protocol, recombinant vitronectin and synthemax® could be used together with commercially available, fully defined, xeno-free essential 8 medium without significantly impacting the reprogramming efficiency. To demonstrate the robustness of this protocol, we reprogrammed a further 2 primary patient cell lines, one with retinosa pigmentosa and the other with Parkinsons disease. We believe that we have optimised a simple and reproducible method which could be used as a starting point for developing GMP protocols, a prerequisite for generating clinically relevant patient specific iPS cells.
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影响因子：
14.8
作者：
通讯作者：
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NuRD 阻止小鼠体细胞重编程为多能干细胞
影响因子：
5.2
作者：
Luo, Min;Ling, Te;Grummt, Ingrid
通讯作者：
Grummt, Ingrid
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.
影响因子：
56.9
作者：
Thomson, JA;Itskovitz-Eldor, J;Jones, JM
通讯作者：
Jones, JM