Linking incomplete reprogramming to the improved pluripotency of murine embryonal carcinoma cell-derived pluripotent stem cells

• Published in: PloS one 2010-04, Vol.5 (4), p.e10320-e10320
• Main Authors: Chang, Gang, Miao, Yi-Liang, Zhang, Yu, Liu, Sheng, Kou, Zhaohui, Ding, Junjun, Chen, Da-Yuan, Sun, Qing-Yuan, Gao, Shaorong
• Format: Article
• Language: English
• Subjects: Analysis; Animals; Biology; Cancer; Carcinoma; Cell Nucleus; Cellular Reprogramming; Cloning; Collaboration; Deoxyribonucleic acid; Developmental Biology; Developmental Biology/Embryology; Developmental Biology/Stem Cells; DNA; DNA binding proteins; DNA Methylation; Embryonal Carcinoma Stem Cells - cytology; Embryonal Carcinoma Stem Cells - ultrastructure; Embryos; Epigenetics; Female; Gene expression; Genes; Genomes; Genomics; H19 gene; Induced Pluripotent Stem Cells - cytology; Induced Pluripotent Stem Cells - metabolism; Intermediates; Laboratories; Leukemia; Methylation; Mice; Molecular modelling; Nuclear transfer; Nuclei; Nuclei (cytology); Oocytes; Overexpression; Pluripotency; Pluripotent Stem Cells - cytology; Pluripotent Stem Cells - metabolism; Somatic cell nuclear transfer; Somatic cells; Stem cell transplantation; Stem cells; Transcription factors; Transcriptional Activation; Tumorigenesis; Tumors; Zoology
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0010320

Somatic cell nuclear transfer (SCNT) has been proved capable of reprogramming various differentiated somatic cells into pluripotent stem cells. Recently, induced pluripotent stem cells (iPS) have been successfully derived from mouse and human somatic cells by the over-expression of a combination of transcription factors. However, the molecular mechanisms underlying the reprogramming mediated by either the SCNT or iPS approach are poorly understood. Increasing evidence indicates that many tumor pathways play roles in the derivation of iPS cells. Embryonal carcinoma (EC) cells have the characteristics of both stem cells and cancer cells and thus they might be the better candidates for elucidating the details of the reprogramming process. Although previous studies indicate that EC cells cannot be reprogrammed into real pluripotent stem cells, the reasons for this remain unclear. Here, nuclei from mouse EC cells (P19) were transplanted into enucleated oocytes and pluripotent stem cells (P19 NTES cells) were subsequently established. Interestingly, P19 NTES cells prolonged the development of tetraploid aggregated embryos compared to EC cells alone. More importantly, we found that the expression recovery of the imprinted H19 gene was dependent on the methylation state in the differential methylation region (DMR). The induction of Nanog expression, however, was independent of the promoter region DNA methylation state in P19 NTES cells. A whole-genome transcriptome analysis further demonstrated that P19 NTES cells were indeed the intermediates between P19 cells and ES cells and many interesting genes were uncovered that may be responsible for the failed reprogramming of P19 cells. To our knowledge, for the first time, we linked incomplete reprogramming to the improved pluripotency of EC cell-derived pluripotent stem cells. The candidate genes we discovered may be useful not only for understanding the mechanisms of reprogramming, but also for deciphering the transition between tumorigenesis and pluripotency.