Rapid and efficient reprogramming of human amnion-derived cells into pluripotency by three factors OCT4/SOX2/NANOG.
Reprogramming human somatic cells to pluripotency represents a valuable resource for research aiming at the development of in vitro models for human diseases and regenerative medicines to produce patient-specific induced pluripotent stem (iPS) cells. Seeking appropriate cell resources for higher efficiency and reducing the risk of viral transgene activation, especially oncogene activation, are of significance for iPS cell research. In this study, we tested whether human amnion-derived cells (hADCs) could be rapidly and efficiently reprogrammed into iPS cells by the defined factors: OCT4/SOX2/NANOG. hADCs from normal placenta were isolated and cultured. The 3rd passage cells were infected with the lentiviral vectors for the delivery of OCT4, SOX2, and NANOG. Afterwards, the generated iPSCs were identified by morphology, pluripotency markers, global gene expression profiles, and epigenetic status both in vitro and in vivo. The results showed that we were able to reprogram hADCs by the defined factors (OCT4/SOX2/NANOG). The efficiency was significantly high (about 0.1%), and the typical colonies appeared on the 9th day after infection. They were similar to human embryonic stem (ES) cells in morphology, proliferation, surface markers, gene expression, and the epigenetic status of pluripotent cell-specific genes. Furthermore, these cells were able to differentiate into various cell types of all three germ layers both in vitro and in vivo. These results demonstrate that hADCs were an ideal somatic cell resource for the rapid and efficient generation of iPS cells by OCT4/SOX2/NANOG.
Zhao
Hong-xi
HX
Department of Gynecology and Obstetrics, Tangdu Hospital, The Fourth Military Medical University, Xi'an 710038, China. dragon.zhaohx@gmail.com
Li
Yang
Y
Jin
Hai-feng
HF
Xie
Li
L
Liu
Chuang
C
Jiang
Feng
F
Luo
Ya-ning
YN
Yin
Guo-wu
GW
Li
Yi
Y
Wang
Jun
J
Li
Ling-song
LS
Yao
Yuan-qing
YQ
Wang
Xiao-hong
XH
D000469:Alkaline Phosphatase / Q000378:metabolism; D000650:Amnion / Q000166:cytology*; D001483:Base Sequence; D015153:Blotting, Western; D002454:Cell Differentiation / Q000502:physiology*; D002460:Cell Line; D017931:DNA Primers; D005455:Fluorescent Antibody Technique; D020869:Gene Expression Profiling; D018398:Homeodomain Proteins / Q000235:genetics / Q000502:physiology*; D006801:Humans; D000071317:Nanog Homeobox Protein; D050814:Octamer Transcription Factor-3 / Q000235:genetics / Q000502:physiology*; D020411:Oligonucleotide Array Sequence Analysis; D039904:Pluripotent Stem Cells / Q000166:cytology*; D016133:Polymerase Chain Reaction; D055748:SOXB1 Transcription Factors / Q000235:genetics / Q000502:physiology*
D017931:DNA Primers; D018398:Homeodomain Proteins; C486780:NANOG protein, human; D000071317:Nanog Homeobox Protein; D050814:Octamer Transcription Factor-3; C494173:POU5F1 protein, human; C527782:SOX2 protein, human; D055748:SOXB1 Transcription Factors; D000469:Alkaline Phosphatase