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Asymmetricity Between Sister Cells of Pluripotent Stem Cells at the Onset of Differentiation

Various somatic stem cells divide asymmetrically; however, it is not known whether embryonic stem cells (ESCs) divide symmetrically or asymmetrically, not only while maintaining an undifferentiated state but also at the onset of differentiation. In this study, we observed single ESCs using time-laps...

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Autores principales: Nakamura, Shogo, Maruyama, Atsushi, Kondo, Yuki, Kano, Ayumu, De Sousa, Olga M., Iwahashi, Masahiro, Hexig, Bayar, Akaike, Toshihiro, Li, Jingyue, Hayashi, Yohei, Ohnuma, Kiyoshi
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Mary Ann Liebert, Inc. 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5833898/
https://www.ncbi.nlm.nih.gov/pubmed/29336219
http://dx.doi.org/10.1089/scd.2017.0113
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author Nakamura, Shogo
Maruyama, Atsushi
Kondo, Yuki
Kano, Ayumu
De Sousa, Olga M.
Iwahashi, Masahiro
Hexig, Bayar
Akaike, Toshihiro
Li, Jingyue
Hayashi, Yohei
Ohnuma, Kiyoshi
author_facet Nakamura, Shogo
Maruyama, Atsushi
Kondo, Yuki
Kano, Ayumu
De Sousa, Olga M.
Iwahashi, Masahiro
Hexig, Bayar
Akaike, Toshihiro
Li, Jingyue
Hayashi, Yohei
Ohnuma, Kiyoshi
author_sort Nakamura, Shogo
collection PubMed
description Various somatic stem cells divide asymmetrically; however, it is not known whether embryonic stem cells (ESCs) divide symmetrically or asymmetrically, not only while maintaining an undifferentiated state but also at the onset of differentiation. In this study, we observed single ESCs using time-lapse imaging and compared sister cell pairs derived from the same mother cell in either the maintenance or differentiation medium. Mouse ESCs were cultured on E-cadherin-coated glass-based dishes, which allowed us to trace single cells. The undifferentiated cell state was detected by green fluorescent protein (GFP) expression driven by the Nanog promoter, which is active only in undifferentiated cells. Cell population analysis using flow cytometry showed that the peak width indicating distribution of GFP expression broadened when cells were transferred to the differentiation medium compared to when they were in the maintenance medium. This finding suggested that the population of ESCs became more heterogeneous at the onset of differentiation. Using single-cell analysis by time-lapse imaging, we found that although the total survival ratio decreased by changing to differentiation medium, the one-live-one-dead ratio of sister cell pairs was smaller compared with randomly chosen non-sister cell pairs, defined as an unsynchronized cell pair control, in both media. This result suggested that sister cell pairs were more positively synchronized with each other compared to non-sister cell pairs. The differences in interdivision time (the time interval between mother cell division and the subsequent cell division) between sister cells was smaller than that between non-sister cell pairs in both media, suggesting that sister cells divided synchronously. Although the difference in Nanog-GFP intensity between sister cells was smaller than that between non-sister cells in the maintenance medium, it was the same in differentiation medium, suggesting asymmetrical Nanog-GFP intensity. These data suggested that ESCs may divide asymmetrically at the onset of differentiation resulting in heterogeneity.
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spelling pubmed-58338982018-04-11 Asymmetricity Between Sister Cells of Pluripotent Stem Cells at the Onset of Differentiation Nakamura, Shogo Maruyama, Atsushi Kondo, Yuki Kano, Ayumu De Sousa, Olga M. Iwahashi, Masahiro Hexig, Bayar Akaike, Toshihiro Li, Jingyue Hayashi, Yohei Ohnuma, Kiyoshi Stem Cells Dev Original Research Reports Various somatic stem cells divide asymmetrically; however, it is not known whether embryonic stem cells (ESCs) divide symmetrically or asymmetrically, not only while maintaining an undifferentiated state but also at the onset of differentiation. In this study, we observed single ESCs using time-lapse imaging and compared sister cell pairs derived from the same mother cell in either the maintenance or differentiation medium. Mouse ESCs were cultured on E-cadherin-coated glass-based dishes, which allowed us to trace single cells. The undifferentiated cell state was detected by green fluorescent protein (GFP) expression driven by the Nanog promoter, which is active only in undifferentiated cells. Cell population analysis using flow cytometry showed that the peak width indicating distribution of GFP expression broadened when cells were transferred to the differentiation medium compared to when they were in the maintenance medium. This finding suggested that the population of ESCs became more heterogeneous at the onset of differentiation. Using single-cell analysis by time-lapse imaging, we found that although the total survival ratio decreased by changing to differentiation medium, the one-live-one-dead ratio of sister cell pairs was smaller compared with randomly chosen non-sister cell pairs, defined as an unsynchronized cell pair control, in both media. This result suggested that sister cell pairs were more positively synchronized with each other compared to non-sister cell pairs. The differences in interdivision time (the time interval between mother cell division and the subsequent cell division) between sister cells was smaller than that between non-sister cell pairs in both media, suggesting that sister cells divided synchronously. Although the difference in Nanog-GFP intensity between sister cells was smaller than that between non-sister cells in the maintenance medium, it was the same in differentiation medium, suggesting asymmetrical Nanog-GFP intensity. These data suggested that ESCs may divide asymmetrically at the onset of differentiation resulting in heterogeneity. Mary Ann Liebert, Inc. 2018-03-01 2018-03-01 /pmc/articles/PMC5833898/ /pubmed/29336219 http://dx.doi.org/10.1089/scd.2017.0113 Text en © Shogo Nakamura et al. 2018; Published by Mary Ann Liebert, Inc. This Open Access article is distributed under the terms of the Creative Commons License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Original Research Reports
Nakamura, Shogo
Maruyama, Atsushi
Kondo, Yuki
Kano, Ayumu
De Sousa, Olga M.
Iwahashi, Masahiro
Hexig, Bayar
Akaike, Toshihiro
Li, Jingyue
Hayashi, Yohei
Ohnuma, Kiyoshi
Asymmetricity Between Sister Cells of Pluripotent Stem Cells at the Onset of Differentiation
title Asymmetricity Between Sister Cells of Pluripotent Stem Cells at the Onset of Differentiation
title_full Asymmetricity Between Sister Cells of Pluripotent Stem Cells at the Onset of Differentiation
title_fullStr Asymmetricity Between Sister Cells of Pluripotent Stem Cells at the Onset of Differentiation
title_full_unstemmed Asymmetricity Between Sister Cells of Pluripotent Stem Cells at the Onset of Differentiation
title_short Asymmetricity Between Sister Cells of Pluripotent Stem Cells at the Onset of Differentiation
title_sort asymmetricity between sister cells of pluripotent stem cells at the onset of differentiation
topic Original Research Reports
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5833898/
https://www.ncbi.nlm.nih.gov/pubmed/29336219
http://dx.doi.org/10.1089/scd.2017.0113
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