RRC ID 64642
著者 Joshi R, Buchanan JC, Paruchuri S, Morris N, Tavana H.
タイトル Molecular Analysis of Stromal Cells-Induced Neural Differentiation of Mouse Embryonic Stem Cells.
ジャーナル PLoS One
Abstract Deriving specific neural cells from embryonic stem cells (ESCs) is a promising approach for cell replacement therapies of neurodegenerative diseases. When co-cultured with certain stromal cells, mouse ESCs (mESCs) differentiate efficiently to neural cells. In this study, a comprehensive gene and protein expression analysis of differentiating mESCs is performed over a two-week culture period to track temporal progression of cells from a pluripotent state to specific terminally-differentiated neural cells such as neurons, astrocytes, and oligodendrocytes. Expression levels of 26 genes consisting of marker genes for pluripotency, neural progenitors, and specific neuronal, astroglial, and oligodendrocytic cells are tracked using real time q-PCR. The time-course gene expression analysis of differentiating mESCs is combined with the hierarchal clustering and functional principal component analysis (FPCA) to elucidate the evolution of specific neural cells from mESCs at a molecular level. These statistical analyses identify three major gene clusters representing distinct phases of transition of stem cells from a pluripotent state to a terminally-differentiated neuronal or glial state. Temporal protein expression studies using immunohistochemistry demonstrate the generation of neural stem/progenitor cells and specific neural lineages and show a close agreement with the gene expression profiles of selected markers. Importantly, parallel gene and protein expression analysis elucidates long-term stability of certain proteins compared to those with a quick turnover. Describing the molecular regulation of neural cells commitment of mESCs due to stromal signaling will help identify major promoters of differentiation into specific cell types for use in cell replacement therapy applications.
巻・号 11(11)
ページ e0166316
公開日 2016-1-1
DOI 10.1371/journal.pone.0166316
PII PONE-D-16-26567
PMID 27832161
PMC PMC5104328
MeSH Animals Astrocytes / cytology* Astrocytes / metabolism Cell Line Cell Proliferation Coculture Techniques Gene Expression Regulation, Developmental Mice Mouse Embryonic Stem Cells / cytology* Mouse Embryonic Stem Cells / metabolism Neurogenesis* Neurons / cytology* Neurons / metabolism Oligodendroglia / cytology* Oligodendroglia / metabolism Stromal Cells / cytology* Stromal Cells / metabolism Transcriptome
IF 2.74
リソース情報
ヒト・動物細胞 MC3T3-G2/PA6(RCB1127)