Characterization of bovine embryos cultured under conditions appropriate for sustaining human naïve pluripotency
In mammalian preimplantation development, pluripotent cells are set aside from cells that contribute to extra-embryonic tissues. Although the pluripotent cell population of mouse and human embryos can be cultured as embryonic stem cells, little is known about the pathways involved in formation of a...
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Published in | PloS one Vol. 12; no. 2; p. e0172920 |
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Main Authors | , , , , , |
Format | Journal Article |
Language | English |
Published |
United States
Public Library of Science
27.02.2017
Public Library of Science (PLoS) |
Subjects | |
Online Access | Get full text |
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Summary: | In mammalian preimplantation development, pluripotent cells are set aside from cells that contribute to extra-embryonic tissues. Although the pluripotent cell population of mouse and human embryos can be cultured as embryonic stem cells, little is known about the pathways involved in formation of a bovine pluripotent cell population, nor how to maintain these cells in vitro. The objective of this study was to determine the transcriptomic profile related to bovine pluripotency. Therefore, in vitro derived embryos were cultured in various culture media that recently have been reported capable of maintaining the naïve pluripotent state of human embryonic cells. Gene expression profiles of embryos cultured in these media were compared using microarray analysis and quantitative RT-PCR. Compared to standard culture conditions, embryo culture in 'naïve' media reduced mRNA expression levels of the key pluripotency markers NANOG and POU5F1. A relatively high percentage of genes with differential expression levels were located on the X-chromosome. In addition, reduced XIST expression was detected in embryos cultured in naïve media and female embryos contained fewer cells with H3K27me3 foci, indicating a delay in X-chromosome inactivation. Whole embryos cultured in one of the media, 5iLA, could be maintained until 23 days post fertilization. Together these data indicate that 'naïve' conditions do not lead to altered expression of known genes involved in pluripotency. Interestingly, X-chromosome inactivation and development of bovine embryos were dependent on the culture conditions. |
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Bibliography: | ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 Conceptualization: BB HH BR.Data curation: BB HvT MGK RW.Formal analysis: BB MGK.Funding acquisition: HH BR.Investigation: BB HvT MGK.Methodology: BB HH BR.Project administration: BB BR.Resources: HvT MGK RW BR.Software: MGK RW.Supervision: HH BR.Validation: BB HH BR.Visualization: BB MGK RW.Writing – original draft: BB MGK RW HH BR.Writing – review & editing: BB HvT MGK RW HH BR. Competing Interests: The authors have declared that no competing interests exist. Current address: Princess Máxima Center for Pediatric Oncology, Utrecht, The Netherlands |
ISSN: | 1932-6203 1932-6203 |
DOI: | 10.1371/journal.pone.0172920 |