机构：[1]Stem Cell Program, Institute for Cell Engineering[2]Graduate Program in Cellular and Molecular Medicine[3]Graduate Program in Biomedical Engineering[4]Division of Hematology in Department of Medicine, The Johns Hopkins University School of Medicine, Baltimore, Maryland, USA[5]Cell Therapy Center, Xuanwu Hospital and Capital Medical University, Beijing, China首都医科大学宣武医院[6]All-Cells LLC, Emeryville, California, USA
To identify accessible and permissive human cell types for efficient derivation of induced pluripotent stem cells (iPSCs), we investigated epigenetic and gene expression signatures of multiple postnatal cell types such as fibroblasts and blood cells. Our analysis suggested that newborn cord blood (CB) and adult peripheral blood (PB) mononuclear cells (MNCs) display unique signatures that are closer to iPSCs and human embryonic stem cells (ESCs) than agematched fibroblasts to iPSCs/ESCs, thus making blood MNCs an attractive cell choice for the generation of integration-free iPSCs. Using an improved EBNA1/OriP plasmid expressing 5 reprogramming factors, we demonstrated highly efficient reprogramming of briefly cultured blood MNCs. Within 14 days of one-time transfection by one plasmid, up to 1000 iPSC-like colonies per 2 million transfected CB MNCs were generated. The efficiency of deriving iPSCs from adult PB MNCs was approximately 50-fold lower, but could be enhanced by inclusion of a second EBNA1/OriP plasmid for transient expression of additional genes such as SV40 T antigen. The duration of obtaining bona fide iPSC colonies from adult PB MNCs was reduced to half (similar to 14 days) as compared to adult fibroblastic cells (2830 days). More than 9 human iPSC lines derived from PB or CB blood cells are extensively characterized, including those from PB MNCs of an adult patient with sickle cell disease. They lack V(D)J DNA rearrangements and vector DNA after expansion for 10-12 passages. This facile method of generating integration-free human iPSCs from blood MNCs will accelerate their use in both research and future clinical applications.
基金：
Johns Hopkins University, a New York State Stem Cell research grant (N08T-019), and grants from the NIH (R01HL073781, U01HL099775, RC2HL101582 and T32HL007525).
第一作者机构：[1]Stem Cell Program, Institute for Cell Engineering
共同第一作者：
通讯作者：
通讯机构：[*]The Johns Hopkins University School of Medicine, Broadway Research Building, Room 747, 733 N. Broadway, Baltimore, MD 21205
推荐引用方式(GB/T 7714)：
Bin-Kuan Chou,Prashant Mali,Xiaosong Huang,et al.Efficient human iPS cell derivation by a non-integrating plasmid from blood cells with unique epigenetic and gene expression signatures[J].CELL RESEARCH.2011,21(3):518-529.doi:10.1038/cr.2011.12.
APA：
Bin-Kuan Chou,Prashant Mali,Xiaosong Huang,Zhaohui Ye,Sarah N Dowey...&Linzhao Cheng.(2011).Efficient human iPS cell derivation by a non-integrating plasmid from blood cells with unique epigenetic and gene expression signatures.CELL RESEARCH,21,(3)
MLA：
Bin-Kuan Chou,et al."Efficient human iPS cell derivation by a non-integrating plasmid from blood cells with unique epigenetic and gene expression signatures".CELL RESEARCH 21..3(2011):518-529
生物学