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Generation of Integration-free Induced Pluripotent Stem Cells from Human Peripheral Blood Mononuclear Cells Using Episomal Vectors
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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
Chapters
- 00:05Title
- 01:01Isolation of Peripheral Blood Mononuclear Cells (PB MNCs)
- 03:03Expansion of PB MNCs in Erythroid Medium
- 04:07PB MNCs Nucleofection and Reprogramming
- 08:00Results: Generation of Integration-free iPSCs from PB MNCs
- 08:48Conclusion
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.
