title: Characterization of bovine embryos cultured under conditions appropriate for sustaining human naïve pluripotency.
abstract: 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.
authors.affiliation: Department of Farm Animal Health, Faculty of Veterinary Medicine, Utrecht University, Utrecht, The Netherlands.
authors.forename: Bas
authors.identifier: http://orcid.org/0000-0003-3617-0709
authors.initials: B
authors.lastname: Brinkhof
authors.affiliation: Department of Farm Animal Health, Faculty of Veterinary Medicine, Utrecht University, Utrecht, The Netherlands.
authors.forename: Helena T A
authors.initials: HT
authors.lastname: van Tol
authors.affiliation: Molecular Cancer Research, University Medical Center Utrecht, Utrecht, The Netherlands.
authors.forename: Marian J A
authors.initials: MJ
authors.lastname: Groot Koerkamp
authors.affiliation: Center for Cellular Imaging (CCI), Department of Biochemistry and Cell Biology, Faculty of Veterinary Medicine, Utrecht University, Utrecht, The Netherlands.
authors.forename: Richard W
authors.initials: RW
authors.lastname: Wubbolts
authors.affiliation: Department of Infectious Diseases and Immunology, Faculty of Veterinary Medicine, Utrecht University, Utrecht, The Netherlands.
authors.forename: Henk P
authors.initials: HP
authors.lastname: Haagsman
authors.affiliation: Department of Farm Animal Health, Faculty of Veterinary Medicine, Utrecht University, Utrecht, The Netherlands.
authors.forename: Bernard A J
authors.initials: BA
authors.lastname: Roelen
mesh_terms: D000818:Animals; D001755:Blastocyst / Q000166:cytology; D002417:Cattle; D003470:Culture Media / Q000378:metabolism; D046149:Embryo Culture Techniques; D047108:Embryonic Development; D053595:Embryonic Stem Cells / Q000166:cytology*; D005260:Female; D020869:Gene Expression Profiling; D018507:Gene Expression Regulation, Developmental; D005801:Genes, Homeobox; D006801:Humans; D056484:Jumonji Domain-Containing Histone Demethylases / Q000378:metabolism; D051379:Mice; D000071317:Nanog Homeobox Protein / Q000378:metabolism; D050814:Octamer Transcription Factor-3 / Q000378:metabolism; D020411:Oligonucleotide Array Sequence Analysis; D039904:Pluripotent Stem Cells / Q000166:cytology*; D012333:RNA, Messenger / Q000378:metabolism; D059467:Transcriptome
chemical_list: D003470:Culture Media; D000071317:Nanog Homeobox Protein; D050814:Octamer Transcription Factor-3; C494173:POU5F1 protein, human; D012333:RNA, Messenger; D056484:Jumonji Domain-Containing Histone Demethylases; C524220:KDM6B protein, human