Bidirectional reprogramming of mouse embryonic stem cell/fibroblast hybrid cells is initiated at the heterokaryon stage

Gridina, Maria; Serov, O. L.

doi:10.1007/s00441-010-1085-2

Cited by 11 publications

(12 citation statements)

References 27 publications

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“…Experiments with hybrid cells conducted by Blau et al 25 suggested the possibility of broad epigenetic changes at a quiescent heterokaryon stage. In agreement with this, we 26 and others 27 have shown that the molecular changes that determine the success and direction (or dominance) of reprogramming occur during the first hours after cell fusion, even before the first cell division.…”

Section: Discussionsupporting

confidence: 88%

Cell divisions are not essential for the direct conversion of fibroblasts into neuronal cells

et al. 2015

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These authors equally contributed to this work.Keywords: cell division, direct conversion, fibroblast, neuron, reprogramming, transdifferentiation Abbreviations: iPS cells, induced pluripotent stem cells; ES cells, embryonic stem cells; BAM, Brn2, Ascl1 and Myt1l; BAMCM, Brn2, Ascl1, Myt1l and cMyc; DOX, doxycycline; BrdU, bromodeoxyuridine; MEF, mouse embryonic fibroblasts.Direct lineage conversion is a promising approach for disease modeling and regenerative medicine. Cell divisions play a key role in reprogramming of somatic cells to pluripotency, however their role in direct lineage conversion is not clear. Here we used transdifferentiation of fibroblasts into neuronal cells by forced expression of defined transcription factors as a model system to study the role of cellular division in the direct conversion process. We have shown that conversion occurs in the presence of the cell cycle inhibitors aphidicolin or mimosine. Moreover, overexpression of the cell cycle activator cMyc negatively influences the process of direct conversion. Overall, our results suggest that cell divisions are not essential for the direct conversion of fibroblasts into neuronal cells.

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Section: Discussionsupporting

confidence: 88%

Cell divisions are not essential for the direct conversion of fibroblasts into neuronal cells

et al. 2015

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“…Immunofluorescence analysis was carried out according to previously published protocol [ 47 ]. In short, cells cultured on glass coverslips were fixed with 3% formaldehyde (Fluka, Germany) and permeabilized with 0.1% Triton X-100 (Fluka, Germany).…”

Section: Methodsmentioning

confidence: 99%

Comparison of American mink embryonic stem and induced pluripotent stem cell transcriptomes

et al. 2015

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BackgroundRecently fibroblasts of many mammalian species have been reprogrammed to pluripotent state using overexpression of several transcription factors. This technology allows production of induced pluripotent stem (iPS) cells with properties similar to embryonic stem (ES) cells. The completeness of reprogramming process is well studied in such species as mouse and human but there is not enough data on other species. We produced American mink (Neovison vison) ES and iPS cells and compared these cells using transcriptome analysis.ResultsWe report the generation of 10 mink ES and 22 iPS cell lines. The majority of the analyzed cell lines had normal diploid chromosome number. The only ES cell line with XX chromosome set had both X-chromosomes in active state that is characteristic of pluripotent cells. The pluripotency of ES and iPS cell lines was confirmed by formation of teratomas with cell types representing all three germ layers. Transcriptome analysis of mink embryonic fibroblasts (EF), two ES and two iPS cell lines allowed us to identify 11831 assembled contigs which were annotated. These led to a number of 6891 unique genes. Of these 3201 were differentially expressed between mink EF and ES cells. We analyzed expression levels of these genes in iPS cell lines. This allowed us to show that 80% of genes were correctly reprogrammed in iPS cells, whereas approximately 6% had an intermediate expression pattern, about 7% were not reprogrammed and about 5% had a "novel" expression pattern. We observed expression of pluripotency marker genes such as Oct4, Sox2 and Rex1 in ES and iPS cell lines with notable exception of Nanog.ConclusionsWe had produced and characterized American mink ES and iPS cells. These cells were pluripotent by a number of criteria and iPS cells exhibited effective reprogramming. Interestingly, we had showed lack of Nanog expression and consider it as a species-specific feature.

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“…All three cell types are reasonably pluripotent as judged by EB formation (this study) and other research (Ambrosi et al, 2007;Battulin et al, 2012;Gridina and Serov, 2010). FMRCs performed well in EB differentiation, but are not as pluripotent as the other two cell types because FMRCs cannot yield mice by tetraploid embryo complementation, although they can contribute to chimeras (Battulin et al, 2012).…”

mentioning

confidence: 91%

Oct4Promoter Activity in Stem Cells Obtained through Somatic Reprogramming

Krueger

Tanasijevic

Norris

et al. 2013

Cellular Reprogramming

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Multiple methods exist that can reprogram differentiated cells to a pluripotent state similar to that of embryonic stem cells (ESCs). These include somatic cell nuclear transfer (SCNT), fusion-mediated reprogramming (FMR) of somatic cells with ESCs, and the production of induced pluripotent stem cells (iPSCs). All of these methods yield cells in which the endogenous Oct4 gene is reactivated. We were interested in comparing the activity of the Oct4 promoter in three different classes of pluripotent cells, including normal ESCs, FMR cells (FMRCs), and iPSCs. We prepared cells of all three types that harbor a transgene composed of the mouse Oct4 promoter driving green fluorescent protein (Oct4-GFP). All cell derivations started with a characterized transgenic Oct4-GFP mouse, and from this we derived ESCs, FMRCs, and iPSCs with the Oct4-GFP transgene present in an identical genomic integration site in all three cell types. Using flow cytometry we assessed Oct4 promoter expression, cell cycle behavior, and differentiation kinetics. We found similar levels of GFP expression in all three cell types and no significant alterations in pluripotency or differentiation. Our results suggest that the pluripotent condition is a potent "local attractor" state, because it can be achieved through three vastly different avenues.

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Bidirectional reprogramming of mouse embryonic stem cell/fibroblast hybrid cells is initiated at the heterokaryon stage

Cited by 11 publications

References 27 publications

Cell divisions are not essential for the direct conversion of fibroblasts into neuronal cells

Cell divisions are not essential for the direct conversion of fibroblasts into neuronal cells

Comparison of American mink embryonic stem and induced pluripotent stem cell transcriptomes

Oct4Promoter Activity in Stem Cells Obtained through Somatic Reprogramming

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