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- Title
Comparative Proteomic Analysis of Human Somatic Cells, Induced Pluripotent Stem Cells, and Embryonic Stem Cells.
- Authors
S. Y. Kim; M. J. Kim; M. J. Son; Y. M. Han; S. Ch. Lee; Y. S. Cho; K. H. Bae
- Abstract
Objective: The purpose of this study was to perform a detailed analysis of the proteomes of somatic donor cells, human iPSCs (hiPSCs) derived from the corresponding somatic cells, and human ESCs (hESCs) to validate the usefulness of hiPSCs at the proteomic level. Materials and Methods: Generation of hiPSCs, Maintenance of hiPSCs and hESCs, Polymerase chain reaction analysis, Immunocytochemistry, Karyotype analysis, Isoelectric focusing and 2-D gel electrophoresis, Staining and image analysis, In-gel digestion and identification by liquid chromatography-MS/MS, Target validation using western blot analysis. Results: Generation of hiPSCs, Proteomic analysis of donor cells, hiPSCs, and hESCs by 2-DE, Identification and classification of differentially expressed proteins by liquid chromotography-MS/MS, Validation of proteins by western blot analysis, Protein network analysis of identified proteins. Conclusion: We performed a comparative proteome analysis of hESCs, hiPSCs, and their corresponding donor cells (hFFs). Through this approach, we identified many proteins that may be directly or indirectly involved in reprogramming. The identified proteins are involved in various biological processes, including transcription cofactor activity, proteasome activator activity, lipid metabolic processes, cell redox homeostasis, and nucleoside metabolic processes, indicating thatsignificant physiological changes occur during reprogramming. Further, we identified several proteins with differential expression patterns between hESCs and hiPSCs. In future studies, we will perform a detailed investigation of the roles of the identified proteins during reprogramming and examine whether they can be effectively utilized to induce or regulate reprogramming at will. In combination with our proteomic analyses, further characterization of these proteins should provide valuable new insights into the mechanism of reprogramming
- Subjects
PROTEOMICS; PROTEIN genetics; SOMATIC cells; PLURIPOTENT stem cells; IMMUNOCYTOCHEMISTRY; ELECTROPHORESIS
- Publication
Cell Journal (Yakhteh), 2013, Vol 15, Issue Sup 1, p53
- ISSN
2228-5806
- Publication type
Article