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血液细胞重编程

实现您将疾病模型培养在培养皿中的理想目标

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hiPS细胞建系

专用培养基  订制服务  24h技术支持

Generation of Integration-Free iPS Cells From 5ml Human Blood


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实验流程:

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载体信息:

pEV-SFFV-OCT4-Wpre; 

pEV-SFFV-SOX2-Wpre; 

pEV-SFFV-Myc-Wpre; 

pEV-SFFV-KLF4-Wpre; 

pEV-SFFV-OCT4-E2A-SOX2-Wpre;

pEV-SFFV-MYC-E2A-KLF4-Wpre; 

pEV-SFFV-BCL-XL-Wpre


注:

SFFV: spleen focus-forming virus U3 promoter; 

 E2A: a self-cleavage site derived from equine rhinitis A virus,(CAG TGT ACT AAT TAT GCT CTC TTG AAA TTG GCT GGA GAT GTT GAG AGC AAC CCA GGT CCC); 

WPER: posttranscriptional regulatory element;

SV40PolyA: polyadenylation signal from SV40 virus; 

OriP: EBV origin of replication; 

EBNA1: Epstein–Barr nuclear antigen 1, which plays essential roles in replication and persistence of episomal plasmid in infected cells.


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参考文献:

1. Meng X, Neises A, Su RJ, Payne KJ, Ritter L, Gridley DS, Wang J, Sheng M, Lau KH, Baylink DJ, Zhang XB: Efficient reprogramming of human cord blood CD34+ cells into induced pluripotent stem cells with OCT4 and SOX2 alone. Molecular Therapy, 2012;20(2):408-16.


2. Su RJ, Baylink DJ, Neises A, Kiroyan JB, Meng X, Payne KJ, Tschudy-Seney B, Duan Y, Appleby N, Kearns-Jonker M, Gridley DS, Wang J, Lau KHW, Zhang XB: Efficient generation of integration-free iPS cells from human adult peripheral blood using BCL-XL together with Yamanaka factors. PLoS One. 2013, 8(5): e64496.


3. Zhang XB: Cellular reprogramming of human peripheral blood cells. Genomics Proteomics Bioinformatics. 2013 Oct;11(5):264-74.


4. Su RJ, Neises A, Zhang XB: Generation of iPS cells from human peripheral blood mononuclear cells using episomal vectors. Methods Mol Biol. 2016;1357:57-69.


5. Wen W#, Zhang JP#, Xu J#, Su RJ, Neises A, Ji GZ, Yuan W, Cheng T*, Zhang XB*. Enhanced generation of integration-free iPSCs from human adult peripheral blood mononuclear cells with an optimal combination of episomal vectors. Stem Cell Reports. 2016 Jun 14;6(6):873-84.


6. Wen W, Zhang JP, Chen W, Arakaki C, Li XL, Baylink D, Botimer GD, Xu J, Yuan W, Cheng T*, Zhang XB*. Generation of integration-free induced pluripotent stem cells from human peripheral blood mononuclear cells using episomal vectors. J Vis Exp. 2017 Jan 1;(119). doi: 10.3791/55091.


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