Generation of Integration-free Induced Pluripotent Stem Cells from Human Peripheral Blood Mononuclear Cells Using Episomal Vectors

Wei Wen; Jian-Ping Zhang; Wanqiu Chen; Cameron Arakaki; Xiaolan Li; David Baylink; Gary D. Botimer; Jing Xu; Weiping Yuan; Tao Cheng; Xiao-Bing Zhang

doi:10.3791/55091

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Generazione di integrazione-liberi cellule staminali pluripotenti indotte dalle cellule mononucleari di sangue periferico umano Utilizzando episomiale Vettori

Generation of Integration-free Induced Pluripotent Stem Cells from Human Peripheral Blood Mononuclear Cells Using Episomal Vectors

JoVE Journal
Biologia dello sviluppo

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JoVE Journal Biologia dello sviluppo

Generation of Integration-free Induced Pluripotent Stem Cells from Human Peripheral Blood Mononuclear Cells Using Episomal Vectors

Generazione di integrazione-liberi cellule staminali pluripotenti indotte dalle cellule mononucleari di sangue periferico umano Utilizzando episomiale Vettori

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10,645 Views

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09:45 min

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January 01, 2017

DOI:

10.3791/55091-v

DOI:

10.3791/55091-v

•

09:45 min

January 01, 2017

•

10634 Views

Wei Wen¹, Jian-Ping Zhang¹, Wanqiu Chen², Cameron Arakaki², Xiaolan Li^1,2, David Baylink², Gary D. Botimer³, Jing Xu¹, Weiping Yuan¹, Tao Cheng^1,4,5,6,7, Xiao-Bing Zhang^1,2

¹State Key Laboratory of Experimental Hematology, Institute of Hematology and Blood Disease Hospital,Chinese Academy of Medical Sciences and Peking Union Medical College ²Division of Regenerative Medicine, Department of Medicine,Loma Linda University ³Department of Orthopaedic Surgery,Loma Linda University ⁴Center for Stem Cell Medicine,Chinese Academy of Medical Sciences ⁵Department of Stem Cell & Regenerative Medicine,Peking Union Medical College ⁶Collaborative Innovation Center for Cancer Medicine ⁷Tianjin Key Laboratory of Blood Cell Therapy and Technology

Summary

This protocol describes a detailed method for efficient generation of integration-free iPSCs from human adult peripheral blood cells. With the use of four oriP/EBNA-based episomal vectors to express the reprogramming factors, KLF4, MYC, BCL-XL, or OCT4 and SOX2, thousands of iPSC colonies can be obtained from 1 mL of peripheral blood.