6 results on '"Sevlever GE"'
Search Results
2. miR-302 family, miR-145 and miR-296 temporal expression profile along the cell cycle of human pluripotent stem cells.
- Author
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Rodríguez-Varela MS, Mucci S, Videla-Richardson GA, Isaja L, Sevlever GE, Scassa ME, and Romorini L
- Subjects
- Cell Line, Cytostatic Agents pharmacology, Humans, Induced Pluripotent Stem Cells drug effects, Induced Pluripotent Stem Cells physiology, MicroRNAs metabolism, Cell Cycle Checkpoints, Induced Pluripotent Stem Cells metabolism, MicroRNAs genetics
- Abstract
Human pluripotent stem cells (hPSCs), like embryonic (hESCs) and induced pluripotent stem cells (hiPSCs), exhibit an unusual cell cycle structure characterized by a short G1 phase and cells being most of time in S phase. hPSCs are receptive to differentiation cues during their transition through G1 phase when lineage determination is decided. Although several MicroRNAs (miRNAs) have been shown to target transcripts that directly or indirectly coordinate the cell cycle of pluripotent cells, its temporal expression profile along hPSCs cell cycle remains poorly characterized. miR-145 and miR-296 are induced during differentiation and silence the self-renewal and pluripotency program. miR-302 family is essential for hPSCs stemness and its expression decreases during differentiation. We aimed to study how the aforementioned miRNAs are regulated along the cell cycle of hPSCs. We demonstrated by pharmacological synchronization and block and release experiments that miR-145, miR-296 and miR-302 family are periodically expressed in hPSCs. Importantly, miR-302 family expression is induced at G1/S boundary and remained high at S phase, presumably to impede differentiation onset. Besides, we confirmed by a gene ontology analysis that many validated miR-302 family target genes are involved in cell cycle regulation., (Copyright © 2021 Elsevier B.V. All rights reserved.)
- Published
- 2021
- Full Text
- View/download PDF
3. Generation of a human induced pluripotent stem cell line from a familial Alzheimer's disease PSEN1 T119I patient.
- Author
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Isaja L, Rodríguez-Varela MS, Marazita M, Mucci S, Itzcovich T, Chrem-Méndez P, Niikado M, Ferriol-Laffouillere SL, Allegri R, Martinetto H, Sevlever GE, Scassa ME, Surace EI, and Romorini L
- Subjects
- Aged, Cell Differentiation, Fibroblasts, Humans, Male, Presenilin-1 genetics, Alzheimer Disease genetics, Induced Pluripotent Stem Cells
- Abstract
Human induced pluripotent stem cells (hiPSC) line FLENIi001-A was reprogrammed from dermal fibroblasts using the lentiviral-hSTEMCCA-loxP vector. Fibroblasts were obtained from a skin biopsy of a 72-year-old Caucasian male familial Alzheimer's disease patient carrying the T119I mutation in the PSEN1 gene. PSEN1 genotype was maintained and stemness and pluripotency confirmed in the FLENIi001-A hiPSC line., (Copyright © 2021 The Authors. Published by Elsevier B.V. All rights reserved.)
- Published
- 2021
- Full Text
- View/download PDF
4. MicroRNA characterization in equine induced pluripotent stem cells.
- Author
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Moro LN, Amin G, Furmento V, Waisman A, Garate X, Neiman G, La Greca A, Santín Velazque NL, Luzzani C, Sevlever GE, Vichera G, and Miriuka SG
- Subjects
- Animals, Cell Differentiation genetics, Fibroblasts cytology, Gene Expression Profiling, Induced Pluripotent Stem Cells cytology, Kruppel-Like Factor 4, Nuclear Transfer Techniques, Horses, Induced Pluripotent Stem Cells metabolism, MicroRNAs genetics
- Abstract
Cell reprogramming has been well described in mouse and human cells. The expression of specific microRNAs has demonstrated to be essential for pluripotent maintenance and cell differentiation, but not much information is available in domestic species. We aim to generate horse iPSCs, characterize them and evaluate the expression of different microRNAs (miR-302a,b,c,d, miR-205, miR-145, miR-9, miR-96, miR-125b and miR-296). Two equine iPSC lines (L2 and L3) were characterized after the reprogramming of equine fibroblasts with the four human Yamanaka's factors (OCT-4/SOX-2/c-MYC/KLF4). The pluripotency of both lines was assessed by phosphatase alkaline activity, expression of OCT-4, NANOG and REX1 by RT-PCR, and by immunofluorescence of OCT-4, SOX-2 and c-MYC. In vitro differentiation to embryo bodies (EBs) showed the capacity of the iPSCs to differentiate into ectodermal, endodermal and mesodermal phenotypes. MicroRNA analyses resulted in higher expression of the miR-302 family, miR-9 and miR-96 in L2 and L3 vs. fibroblasts (p<0.05), as previously shown in human pluripotent cells. Moreover, downregulation of miR-145 and miR-205 was observed. After differentiation to EBs, higher expression of miR-96 was observed in the EBs respect to the iPSCs, and also the expression of miR-205 was induced but only in the EB-L2. In addition, in silico alignments of the equine microRNAs with mRNA targets suggested the ability of miR-302 family to regulate cell cycle and epithelial mesenchymal transition genes, miR-9 and miR-96 to regulate neural determinant genes and miR-145 to regulate pluripotent genes, similarly as in humans. In conclusion, we could obtain equine iPSCs, characterize them and determine for the first time the expression level of microRNAs in equine pluripotent cells., Competing Interests: We have the following interests: Author Gabriel Vichera is employed by Kheiron Biotech. There are no patents, products in development or marketed products to declare. This does not alter our adherence to all the PLOS ONE policies on sharing data and materials.
- Published
- 2018
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5. Human Pluripotent Stem Cells and Derived Neuroprogenitors Display Differential Degrees of Susceptibility to BH3 Mimetics ABT-263, WEHI-539 and ABT-199.
- Author
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García CP, Videla Richardson GA, Dimopoulos NA, Fernandez Espinosa DD, Miriuka SG, Sevlever GE, Romorini L, and Scassa ME
- Subjects
- Ataxia Telangiectasia Mutated Proteins metabolism, Camptothecin pharmacology, Cell Line, Human Embryonic Stem Cells cytology, Humans, Induced Pluripotent Stem Cells cytology, Neural Stem Cells cytology, Proto-Oncogene Proteins c-bcl-2 metabolism, Tumor Suppressor Protein p53 metabolism, bcl-X Protein metabolism, Aniline Compounds pharmacology, Bridged Bicyclo Compounds, Heterocyclic pharmacology, Human Embryonic Stem Cells metabolism, Induced Pluripotent Stem Cells metabolism, Neural Stem Cells metabolism, Sulfonamides pharmacology
- Abstract
Human embryonic stem cells (hESCs) are hypersensitive to genotoxic stress and display lower survival ability relative to their differentiated progeny. Herein, we attempted to investigate the source of this difference by comparing the DNA damage responses triggered by the topoisomerase I inhibitor camptothecin, in hESCs, human induced pluripotent stem cells (hiPSCs) and hESCs-derived neuroprogenitors (NP). We observed that upon camptothecin exposure pluripotent stem cells underwent apoptosis more swiftly and at a higher rate than differentiated cells. However, the cellular response encompassing ataxia-telangiectasia mutated kinase activation and p53 phosphorylation both on serine 15 as well as on serine 46 resulted very similar among the aforementioned cell types. Importantly, we observed that hESCs and hiPSCs express lower levels of the anti-apoptotic protein Bcl-2 than NP. To assess whether Bcl-2 abundance could account for this differential response we treated cells with ABT-263, WEHI-539 and ABT-199, small molecules that preferentially target the BH3-binding pocket of Bcl-xL and/or Bcl-2 and reduce their ability to sequester pro-apoptotic factors. We found that in the absence of stress stimuli, NP exhibited a higher sensitivity to ABT- 263 and WEHI-539 than hESCs and hiPSCs. Conversely, all tested cell types appeared to be highly resistant to the Bcl-2 specific inhibitor, ABT-199. However, in all cases we determined that ABT-263 or WEHI-539 treatment exacerbated camptothecin-induced apoptosis. Importantly, similar responses were observed after siRNA-mediated down-regulation of Bcl-xL or Bcl-2. Taken together, our results suggest that Bcl-xL contrary to Bcl-2 contributes to ensure cell survival and also functions as a primary suppressor of DNA double-strand brake induced apoptosis both in pluripotent and derived NP cells. The emerging knowledge of the relative dependence of pluripotent and progenitor cells on Bcl-2 and Bcl-xL activities may help to predict cellular responses and potentially manipulate these cells for therapeutic purposes in the near future.
- Published
- 2016
- Full Text
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6. Generation of iPSC line iPSC-FH2.1 in hypoxic conditions from human foreskin fibroblasts.
- Author
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Questa M, Romorini L, Blüguermann C, Solari CM, Neiman G, Luzzani C, Scassa MÉ, Sevlever GE, Guberman AS, and Miriuka SG
- Subjects
- Cell Differentiation, Cells, Cultured, Cellular Reprogramming, Comparative Genomic Hybridization, DNA Methylation, Humans, Induced Pluripotent Stem Cells metabolism, Karyotype, Kruppel-Like Factor 4, Male, Microscopy, Fluorescence, Octamer Transcription Factor-3 genetics, Promoter Regions, Genetic, Real-Time Polymerase Chain Reaction, Transcription Factors genetics, Transcription Factors metabolism, Fibroblasts cytology, Foreskin cytology, Induced Pluripotent Stem Cells cytology
- Abstract
Human foreskin fibroblasts were used to generate the iPSC line iPSC-FH2.1 using the EF1a-hSTEMCCA-loxP vector expressing OCT4, SOX2, c-MYC and KLF4, in 5% O2 culture conditions. Stemness was confirmed, as was pluripotency both in vivo and in vitro, in normoxia and hypoxia. Human Embryonic Stem Cell (hESC) line WA-09 and reprogrammed fibroblast primary culture HFF-FM were used as controls., (Copyright © 2015 University of Texas at Austin Dell Medical School. Published by Elsevier B.V. All rights reserved.)
- Published
- 2016
- Full Text
- View/download PDF
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