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Pluripotency can be rapidly and efficiently induced in human amniotic fluid-derived cells
Chunliang Li1,4,#, Junmei Zhou5,6,#, Guilai Shi1,4, Yu Ma1,2, Ying Yang1,4, Junjie Gu1,2, Hongyao Yu1,4, Shibo Jin1,2, Zhe Wei1,4, Fang Chen5,6 and Ying Jin1,2,3,*
1 Key Laboratory of Stem Cell Biology, Institute of Health Sciences, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences/Shanghai JiaoTong University School of Medicine; 225 South Chongqing Road, Shanghai, 200025, China 2 Shanghai Stem Cell Institute, Shanghai JiaoTong University School of Medicine, Shanghai, China 3 Key Laboratory of Cell Differentiation and Apoptosis of Chinese Ministry of Education, Shanghai JiaoTong University School of Medicine, Shanghai, China 4 Graduate School of Chinese Academy of Sciences, Beijing, China 5 Department of Urology, Shanghai Children's Hospital, Shanghai JiaoTong University School of Medicine, Shanghai, China 6 Tissue Engineering Laboratory, Xinhua Hospital, Shanghai Institute for Pediatric Research, Shanghai JiaoTong University School of Medicine, Shanghai, China
Direct reprogramming of human somatic cells into pluripotency has broad implications in generating patient-specific induced pluripotent stem (iPS) cells for disease modeling and cellular replacement therapies. However, the low efficiency and safety issues associated with generation of human iPS cells have limited their usage in clinical settings. Cell types can significantly influence reprogramming efficiency and kinetics. To date, human iPS cells have been obtained only from a few cell types. Here, we report for the first time rapid and efficient generation of iPS cells from human amniotic fluid-derived cells (hAFDCs) via ectopic expression of four human factors: OCT4/SOX2/KLF4/C-MYC. Significantly, typical single iPS cell colonies can be picked up six days after viral infection with high efificiency. Eight iPS cell lines have been derived. They can be continuously propagated in vitro and express pluripotency markers such as AKP, OCT4, SOX2, SSEA4, TRA-1-60 and TRA-1-81, maintaining the normal karyotype. Transgenes are completely inactivated and the endogenous OCT4 promoter is adequately demethylated in the established iPS cell lines. Moreover, various cells and tissues from all three germ layers are found in embryoid bodies and teratomas respectively. In addition, microarray analysis demonstrates a high correlation coefficient between hAFDC-iPS cells and human embryonic stem cells, but a low correlation coefficient between hAFDCs and hAFDC-iPS cells. Taken together, these data identify an ideal human somatic cell resource for rapid and efficient generation of iPS cells, allowing us to establish human iPS cells using more advanced approaches and possibly to establish disease- or patient-specific iPS cells.
