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1.
J Assist Reprod Genet ; 37(1): 149-158, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31701304

RESUMO

PURPOSE: To detect putative differences in the miRNomic profile of follicular fluids collected after follicular-phase-stimulation (FPS-FFs) and paired luteal-phase-stimulation (LPS-FFs) in the same ovarian cycles (DuoStim). METHODS: Exploratory study at a private IVF center and University involving FPS-FFs and paired-LPS-FFs collected from 15 reduced ovarian reserve and advanced maternal age women undergoing DuoStim (n = 30 paired samples). The samples were combined in 6 paired pools (5 samples each) and balanced according to maternal age and number of cumulus-oocyte-complexes. Micro-RNAs were isolated and sequenced. Four miRNAs were then selected for further validation on 6 single pairs of FPS-FFs and LPS-FFs by qPCR. RESULTS: Forty-three miRNAs were detected in both FPS-FFs and paired-LPS-FFs after sequencing and no statistically significant differences were reported. Thirty-three KEGG pathways were identified as regulated from the detected miRNAs. Four miRNAs (miR-146b, miR-191, miR-320a, and miR-483) were selected for qPCR validation since consistently expressed in our samples and possibly involved in the regulation/establishment of a healthy follicular environment. Again, no significant differences were reported between FPS-FFs and paired-LPS-FFs, also when the analysis was corrected for maternal age and number of cumulus-oocyte-complexes in generalized linear models. CONCLUSIONS: These data complement the embryological, chromosomal, and clinical evidence of equivalence between FPS and LPS published to date.


Assuntos
Líquido Folicular/metabolismo , Fase Folicular/genética , Infertilidade Feminina/genética , Fase Luteal/genética , Ciclo Menstrual/genética , MicroRNAs/genética , Indução da Ovulação/métodos , Adulto , Feminino , Fase Folicular/metabolismo , Perfilação da Expressão Gênica , Humanos , Fase Luteal/metabolismo
2.
J Cell Mol Med ; 23(8): 5440-5453, 2019 08.
Artigo em Inglês | MEDLINE | ID: mdl-31237115

RESUMO

Although the concepts of somatic cell reprogramming and human-induced pluripotent stem cells (hiPSCs) generation have undergone several analyses to validate the usefulness of these cells in research and clinic, it remains still controversial whether the hiPSCs are equivalent to human embryonic stem cells (hESCs), pointing to the need of further characterization for a more comprehensive understanding of pluripotency. Most of the experimental evidence comes from the transcriptome analysis, while a little is available on protein data, and even less is known about the post-translational modifications. Here, we report a combined strategy of mass spectrometry and gene expression profiling for proteogenomic analysis of reprogrammed and embryonic stem cells. The data obtained through this integrated, multi-"omics" approach indicate that a small, but still significant, number of distinct pathways is enriched in reprogrammed versus embryonic stem cells, supporting the view that pluripotency is an extremely complex, multifaceted phenomenon, with peculiarities that are characteristic of each cell type.


Assuntos
Células-Tronco Embrionárias Humanas/metabolismo , Células-Tronco Pluripotentes Induzidas/metabolismo , Células Cultivadas , Reprogramação Celular/genética , Fibroblastos/metabolismo , Perfilação da Expressão Gênica/métodos , Humanos , Espectrometria de Massas/métodos , Processamento de Proteína Pós-Traducional/genética , Proteogenômica/métodos , Transcriptoma/genética
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