title: Analysis of differential proteomes of induced pluripotent stem cells by protein-based reprogramming of fibroblasts.
abstract: The recent generation of induced pluripotent stem (iPS) cells represents a novel opportunity to complement embryonic stem (ES) cell-based approaches. iPS cells can be generated by viral transduction of specific transcription factors, but there is a potential risk of tumorigenicity by random retroviral integration. We have generated novel iPS (sFB-protein-iPS) cells from murine dermal fibroblasts (FVB-sFB) that have ES cell characteristics, using ES cell-derived cell extracts instead of performing viral transduction. Notably, only cell extracts from an ES cell line (C57-mES) on the C57/BL6 background generated iPS cells in our protocol-not an ES cell line (E14-mES) on the 129 background. Hypothesizing that determining the differences in these 2 mES cell lines will provide vital insight into the reprogramming machinery, we performed proteomic and global gene expression analysis by iTRAQ and mRNA microarray, respectively. We observed that pluripotent ES cells and ES cell extract-derived iPS cells had differential proteomes and global gene expression patterns. Notably, reprogramming-competent C57-mES cells highly expressed proteins that regulate protein synthesis and metabolism, compared with reprogramming-incompetent 129-mES cells, suggesting that there is a threshold that protein synthetic machinery must exceed to initiate reprogramming.
authors.affiliation: Department of Biomedical Sciences, Seoul National University College of Medicine, Seoul, Korea.
authors.forename: Jonghwa
authors.initials: J
authors.lastname: Jin
authors.forename: Yoo-Wook
authors.initials: YW
authors.lastname: Kwon
authors.forename: Jae Seung
authors.initials: JS
authors.lastname: Paek
authors.forename: Hyun-Jai
authors.initials: HJ
authors.lastname: Cho
authors.forename: Jiyoung
authors.initials: J
authors.lastname: Yu
authors.forename: Ji Yoon
authors.initials: JY
authors.lastname: Lee
authors.forename: In-Sun
authors.initials: IS
authors.lastname: Chu
authors.forename: In-Hyun
authors.initials: IH
authors.lastname: Park
authors.forename: Young-Bae
authors.initials: YB
authors.lastname: Park
authors.forename: Hyo-Soo
authors.initials: HS
authors.lastname: Kim
authors.forename: Youngsoo
authors.initials: Y
authors.lastname: Kim
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000032:analysis; D002853:Chromatography, Liquid / Q000379:methods; D016000:Cluster Analysis; D005347:Fibroblasts / Q000737:chemistry* / Q000166:cytology / Q000502:physiology; D057026:Induced Pluripotent Stem Cells / Q000737:chemistry* / Q000166:cytology; D051379:Mice; D008810:Mice, Inbred C57BL; D046228:Microarray Analysis / Q000379:methods; D020543:Proteome / Q000032:analysis*; D012333:RNA, Messenger / Q000032:analysis; D015398:Signal Transduction; D053719:Tandem Mass Spectrometry / Q000379:methods
chemical_list: D015415:Biomarkers; D020543:Proteome; D012333:RNA, Messenger