RA induces the neural-like cells generated from epigenetic modified NIH/3T3 cells.
Recently, differentiated somatic cells had been reprogrammed to pluripotential state in vitro, and various tissue cells had been elicited from those cells. Epigenetic modifications allow differentiated cells to perpetuate the molecular memory needed for the cells to retain their identity. DNA methylation and histone deacetylation are important patterns involved in epigenetic modification, which take critical roles in regulating DNA expression. In this study, we dedifferentiated NIH/3T3 fibroblasts by 5-aza-2-deoxycytidine (5-aza-dC) and Trichstatin A (TSA) combination, and detected gene expression pattern, DNA methylation level, and differentiation potential of reprogrammed cells. As the results, embryonic marker Sox2, klf4, c-Myc and Oct4 were expressed in reprogrammed NIH/3T3 fibroblasts. Total DNA methylation level was significant decreased after the treatment. Moreover, exposure of the reprogrammed cells to all trans-retinoic acid (RA) medium elicited the generation of neuronal class IIIbeta-tubulin-positive, neuron-specific enolase (NSE)-positive, nestin-positive, and neurofilament light chain (NF-L)-positive neural-like cells.
Zhang
Xi-Mei
XM
Department of Histology and Embryology, Harbin Medical University, Harbin, China.
Li
Qiu-Ming
QM
Su
Dong-Ju
DJ
Wang
Ning
N
Shan
Zhi-Yan
ZY
Jin
Lian-Hong
LH
Lei
Lei
L
D000818:Animals; D001374:Azacitidine / Q000031:analogs & derivatives; D002454:Cell Differentiation / Q000502:physiology*; D019175:DNA Methylation; D000077209:Decitabine; D044127:Epigenesis, Genetic / Q000502:physiology*; D005434:Flow Cytometry; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation / Q000502:physiology*; D007150:Immunohistochemistry; D007381:Intermediate Filament Proteins / Q000378:metabolism; D000090062:Kruppel-Like Factor 4; D051741:Kruppel-Like Transcription Factors / Q000378:metabolism; D051379:Mice; D041681:NIH 3T3 Cells; D009419:Nerve Tissue Proteins / Q000378:metabolism; D064231:Nestin; D009474:Neurons / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D050814:Octamer Transcription Factor-3 / Q000378:metabolism; D010751:Phosphopyruvate Hydratase / Q000378:metabolism; D016271:Proto-Oncogene Proteins c-myc / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D055748:SOXB1 Transcription Factors / Q000378:metabolism; D014212:Tretinoin / Q000378:metabolism* / Q000494:pharmacology
D007381:Intermediate Filament Proteins; C000715468:Klf4 protein, mouse; D000090062:Kruppel-Like Factor 4; D051741:Kruppel-Like Transcription Factors; C489428:Myc protein, mouse; D009419:Nerve Tissue Proteins; C577485:Nes protein, mouse; D064231:Nestin; D050814:Octamer Transcription Factor-3; C494174:Pou5f1 protein, mouse; D016271:Proto-Oncogene Proteins c-myc; D055748:SOXB1 Transcription Factors; C527783:Sox2 protein, mouse; D014212:Tretinoin; D000077209:Decitabine; D010751:Phosphopyruvate Hydratase; D001374:Azacitidine