[Frontiers in Bioscience 17, 1659-1668, January 1, 2012]

iPSCs are transcriptionally and post-transcriptionally indistinguishable from fESCs

Yun-feng Wang1, Jian Li 2,3, Ying-zi He 3, Hui-qian Yu1, Yang Li4, Xiao-dong Gu1, Wen Li1, Hua-wei Li1,3 1Department of Otolaryngology, Affiliated Eye and ENT hospital of Fudan University, Shanghai 200031, China, 2 Department of Cardiology, Affiliated Children hospital of Fudan University, Shanghai 201102,China, 3Institutes of Biomedical Sciences of Fudan University, Shanghai 200032, China, 4Kunming Medical School, China

TABLE OF CONTENTS

1. Abstract
2. Introduction
3. Materials and method
3.1. iPSCs, ntESCs, fESCs and MEFs preparation
3.2. RNA isolation, microarray experiment and data analysis
3.3. Quantitative and semiquantitative RT-PCR analysis
3.4. Cell lysates, in-solution digestion, and iTRAQ labeling
3.5. 2D-LC MS/MS
3.6. MS data analysis
3.7. Data processing and statistical analysis
4. Results
4.1. Development potency of stem cell lines
4.2. Transcriptional profiles of iPSCs, ntESCs and fESCs are highly similar
4.3. Similar levels of transcriptional variability in iPSCs, ntESCs and fESCs
4.4. The protein profiles of iPSCs, ntESCs and fESCs are highly similar
5. Discussion
6. Conclusion
7. Acknowledgements
8. References

1. ABSTRACT

Induced pluripotent stem cells (iPSCs) are generated by reprogramming mouse or human somatic cells to a pluripotent state by introducing key transcription factors and have great therapeutic potential. It has been illustrated that the transcriptional and post-transcriptional profiles of nuclear-transferred embryonic stem cells (ntESCs) is identical to those of embryonic stem cells derived from fertilized blastocysts (fESCs). Although iPSCs seem to be indistinguishable from fESCs, the degree of transcriptomic and proteomic similarity among iPSCs, ntESCs, and fESCs has not yet been elucidated completely. To investigate whether iPSCs and fESCs have similar therapeutic potential, we compared mRNA and protein pro?les of mouse iPSC, ntESCs, and matching fESCs lines using microarray technology, iTRAQ method, and bioinformatic analyses. Real-time PCR, two-dimensional LC, and MS/MS analyses were further conducted to study the expression of speci?c transcripts and identify and quantitate 929 proteins. Our results demonstrate that, like ntESCs, the iPSC and matching fESCs lines have very similar transcriptional and protein expression profiles. This is consistent with their similar developmental potential.