کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
2093261 | 1081949 | 2016 | 12 صفحه PDF | دانلود رایگان |
![عکس صفحه اول مقاله: Enhanced Generation of Integration-free iPSCs from Human Adult Peripheral Blood Mononuclear Cells with an Optimal Combination of Episomal Vectors Enhanced Generation of Integration-free iPSCs from Human Adult Peripheral Blood Mononuclear Cells with an Optimal Combination of Episomal Vectors](/preview/png/2093261.png)
• Expression of MYC and KLF4 with two episomal vectors is critical for PB reprogramming
• Optimized episomal vector combination shows an ∼100-fold increase in reprogramming
• This system is comparable with Sendai virus in generating integration-free iPSCs
SummaryWe previously reported the generation of integration-free induced pluripotent stem cells from adult peripheral blood (PB) with an improved episomal vector (EV) system, which uses the spleen focus-forming virus U3 promoter and an extra factor BCL-XL (B). Here we show an ∼100-fold increase in efficiency by optimizing the vector combination. The two most critical factors are: (1) equimolar expression of OCT4 (O) and SOX2 (S), by using a 2A linker; (2) a higher and gradual increase in the MYC (M) to KLF4 (K) ratio during the course of reprogramming, by using two individual vectors to express M and K instead of one. The combination of EV plasmids (OS + M + K + B) is comparable with Sendai virus in reprogramming efficiency but at a fraction of the cost. The generated iPSCs are indistinguishable from those from our previous approach in pluripotency and phenotype. This improvement lays the foundation for broad applications of episomal vectors in PB reprogramming.
Graphical AbstractFigure optionsDownload as PowerPoint slide
Journal: - Volume 6, Issue 6, 14 June 2016, Pages 873–884