title: Nucleolar molecular signature of pluripotent stem cells.
abstract: Induced pluripotent stem cells (iPSC) are generated by reprogramming somatic cells to the pluripotent state. Identification and quantitative characterization of changes in the molecular organization of the cell during the process of cellular reprogramming is valuable for stem cell research and advancement of its therapeutic applications. Here we employ quantitative Raman microspectroscopy and biomolecular component analysis (BCA) for a comparative analysis of the molecular composition of nucleoli in skin fibroblasts and iPSC derived from them. We report that the cultured fibroblasts obtained from different human subjects, share comparable concentrations of proteins, RNA, DNA, and lipids in the molecular composition of nucleoli. The nucleolar molecular environment is drastically changed in the corresponding iPSC. We measured that the transition from skin fibroblasts to iPSC is accompanied by a statistically significant increase in protein concentrations ~1.3-fold, RNA concentrations ~1.3-fold, and DNA concentrations ~1.4-fold, while no statistically significant difference was found for the lipid concentrations. The analysis of molecular vibrations associated with diverse aminoacids and protein conformations indicates that nucleoli of skin fibroblasts contain similar subsets of proteins, with prevalence of tyrosine. In iPSC, we observed a higher signal from tryptophan with an increase in the random coil and α helix protein conformations, indicating changes in the subset of nucleolar proteins during cell reprogramming. At the same time, the concentrations of major types of macromolecules and protein conformations in the nucleoli of iPSC and human embryonic stem cells (hESC) were found to be similar. We discuss these results in the context of nucleolar function and conclude that the nucleolar molecular content is correlated with the cellular differentiation status. The approach described here shows the potential for spectroscopically monitoring changes in macromolecular organization of the cell at different stages of reprogramming.
authors: Institute for Lasers, Photonics and Biophotonics, University at Buffalo, State University of New York, Buffalo, New York 14260, United States.
authors: Artem
authors: A
authors: Pliss
authors: Andrey N
authors: AN
authors: Kuzmin
authors: Aliaksandr V
authors: AV
authors: Kachynski
authors: Houbo
authors: H
authors: Jiang
authors: Zhixing
authors: Z
authors: Hu
authors: Yong
authors: Y
authors: Ren
authors: Jian
authors: J
authors: Feng
authors: Paras N
authors: PN
authors: Prasad
mesh_terms: D002466:Cell Nucleolus / Q000737:chemistry* / Q000235:genetics; D002478:Cells, Cultured; D065150:Cellular Reprogramming; D004247:DNA / Q000032:analysis; D005347:Fibroblasts / Q000737:chemistry* / Q000166:cytology / Q000378:metabolism; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000737:chemistry* / Q000166:cytology / Q000378:metabolism; D008055:Lipids / Q000032:analysis; D009687:Nuclear Proteins / Q000032:analysis; D012313:RNA / Q000032:analysis; D012867:Skin / Q000166:cytology*; D013059:Spectrum Analysis, Raman / Q000379:methods
chemical_list: D008055:Lipids; D009687:Nuclear Proteins; D012313:RNA; D004247:DNA