Enhanced Generation of Integration-free iPSCs from Human Adult Peripheral Blood Mononuclear Cells with an Optimal Combination of Episomal Vectors
Summary: We 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...
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Series: | Stem Cell Reports |
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doaj-37d9ce8c583f44b2b1e808dbb36cb9242020-11-24T21:42:45ZengElsevierStem Cell Reports2213-67112016-06-0166873884Enhanced Generation of Integration-free iPSCs from Human Adult Peripheral Blood Mononuclear Cells with an Optimal Combination of Episomal VectorsWei Wen0Jian-Ping Zhang1Jing Xu2Ruijun Jeanna Su3Amanda Neises4Guang-Zhen Ji5Weiping Yuan6Tao Cheng7Xiao-Bing Zhang8State Key Laboratory of Experimental Hematology, Institute of Hematology and Blood Diseases Hospital, Collaborative Innovation Center for Cancer Medicine, Chinese Academy of Medical Sciences and Peking Union Medical College, Tianjin 300020, ChinaState Key Laboratory of Experimental Hematology, Institute of Hematology and Blood Diseases Hospital, Collaborative Innovation Center for Cancer Medicine, Chinese Academy of Medical Sciences and Peking Union Medical College, Tianjin 300020, ChinaState Key Laboratory of Experimental Hematology, Institute of Hematology and Blood Diseases Hospital, Collaborative Innovation Center for Cancer Medicine, Chinese Academy of Medical Sciences and Peking Union Medical College, Tianjin 300020, ChinaDivision of Regenerative Medicine MC1528B, Department of Medicine, Loma Linda University, 11234 Anderson Street, Loma Linda, CA 92350, USADivision of Regenerative Medicine MC1528B, Department of Medicine, Loma Linda University, 11234 Anderson Street, Loma Linda, CA 92350, USAState Key Laboratory of Experimental Hematology, Institute of Hematology and Blood Diseases Hospital, Collaborative Innovation Center for Cancer Medicine, Chinese Academy of Medical Sciences and Peking Union Medical College, Tianjin 300020, ChinaState Key Laboratory of Experimental Hematology, Institute of Hematology and Blood Diseases Hospital, Collaborative Innovation Center for Cancer Medicine, Chinese Academy of Medical Sciences and Peking Union Medical College, Tianjin 300020, ChinaState Key Laboratory of Experimental Hematology, Institute of Hematology and Blood Diseases Hospital, Collaborative Innovation Center for Cancer Medicine, Chinese Academy of Medical Sciences and Peking Union Medical College, Tianjin 300020, China; Corresponding authorState Key Laboratory of Experimental Hematology, Institute of Hematology and Blood Diseases Hospital, Collaborative Innovation Center for Cancer Medicine, Chinese Academy of Medical Sciences and Peking Union Medical College, Tianjin 300020, China; Division of Regenerative Medicine MC1528B, Department of Medicine, Loma Linda University, 11234 Anderson Street, Loma Linda, CA 92350, USA; Corresponding authorSummary: We 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. : In this article, Zhang, Cheng, and colleagues show that the use of two episomal vectors to express MYC and KLF4 leads to an ∼100-fold increase in reprogramming of human PB MNCs to pluripotency. This improved episomal vector system is comparable with Sendai virus in generating integration-free iPSCs but at a fraction of the cost. Keywords: human induced pluripotent stem cells, peripheral blood mononuclear cells, episomal vectors, sendai viral vectorshttp://www.sciencedirect.com/science/article/pii/S2213671116300303 |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Wei Wen Jian-Ping Zhang Jing Xu Ruijun Jeanna Su Amanda Neises Guang-Zhen Ji Weiping Yuan Tao Cheng Xiao-Bing Zhang |
spellingShingle |
Wei Wen Jian-Ping Zhang Jing Xu Ruijun Jeanna Su Amanda Neises Guang-Zhen Ji Weiping Yuan Tao Cheng Xiao-Bing Zhang Enhanced Generation of Integration-free iPSCs from Human Adult Peripheral Blood Mononuclear Cells with an Optimal Combination of Episomal Vectors Stem Cell Reports |
author_facet |
Wei Wen Jian-Ping Zhang Jing Xu Ruijun Jeanna Su Amanda Neises Guang-Zhen Ji Weiping Yuan Tao Cheng Xiao-Bing Zhang |
author_sort |
Wei Wen |
title |
Enhanced Generation of Integration-free iPSCs from Human Adult Peripheral Blood Mononuclear Cells with an Optimal Combination of Episomal Vectors |
title_short |
Enhanced Generation of Integration-free iPSCs from Human Adult Peripheral Blood Mononuclear Cells with an Optimal Combination of Episomal Vectors |
title_full |
Enhanced Generation of Integration-free iPSCs from Human Adult Peripheral Blood Mononuclear Cells with an Optimal Combination of Episomal Vectors |
title_fullStr |
Enhanced Generation of Integration-free iPSCs from Human Adult Peripheral Blood Mononuclear Cells with an Optimal Combination of Episomal Vectors |
title_full_unstemmed |
Enhanced Generation of Integration-free iPSCs from Human Adult Peripheral Blood Mononuclear Cells with an Optimal Combination of Episomal Vectors |
title_sort |
enhanced generation of integration-free ipscs from human adult peripheral blood mononuclear cells with an optimal combination of episomal vectors |
publisher |
Elsevier |
series |
Stem Cell Reports |
issn |
2213-6711 |
publishDate |
2016-06-01 |
description |
Summary: We 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. : In this article, Zhang, Cheng, and colleagues show that the use of two episomal vectors to express MYC and KLF4 leads to an ∼100-fold increase in reprogramming of human PB MNCs to pluripotency. This improved episomal vector system is comparable with Sendai virus in generating integration-free iPSCs but at a fraction of the cost. Keywords: human induced pluripotent stem cells, peripheral blood mononuclear cells, episomal vectors, sendai viral vectors |
url |
http://www.sciencedirect.com/science/article/pii/S2213671116300303 |
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