Cultured bovine embryo biopsy conserves methylation marks from original embryo

A major limitation of embryo epigenotyping by chromatin immunoprecipitation analysis is the reduced amount of sample available from an embryo biopsy. We developed an in vitro system to expand trophectoderm cells from an embryo biopsy to overcome this limitation. Thiswork analyzes whether expanded trophectoderm (EX) is representative of the trophectoderm (TE) methylation or adaptation to culture has altered its epigenome. We took a small biopsy from the trophectoderm (30-40 cells) of in vitro produced bovine-hatched blastocysts and cultured it on fibronectin-treated plates until we obtained ~4 × 104 cells. The rest of the embryo was allowed to recover its spherical shape and, subsequently, TE and inner cellmass were separated.We examined whether there were DNA methylation differences between TE and EX of three bovine embryos using whole-genome bisulfite sequencing. As a consequence of adaptation to culture, global methylation, including transposable elements, was higher in EX, with 5.3% of quantified regions showing significant methylation differences between TE and EX. Analysis of individual embryos indicated that TE methylation ismore similar to its EX counterpart than to TE from other embryos. Interestingly, these similarly methylated regions are enriched in CpG islands, promoters and transcription units near genes involved in biological processes important for embryo development. Our results indicate that EX is representative of the embryo in terms of DNA methylation, thus providing an informative proxy for embryo epigenotyping. © The Authors 2017. Published by Oxford University Press on behalf of Society for the Study of Reproduction. All rights reserved.

Bibliographic Details

Main Authors:	Fonseca Balvís, N., Garcia-Martinez, S., Pérez Cerezales, Serafín, Ivanova, E., Gómez Redondo, Isabel, Hamdi, M., Rizos Dimitrios, Dimitrios, Coy, P., Kelsey, G., Gutiérrez Adán, Alfonso
Format:	journal article biblioteca
Language:	English
Published:	Oxford University Press 2017
Subjects:	Trophectoderm, Methylation, Whole-genome bisulfite sequencing, Bovine embryo, Epigenotyping, Epigenome, Trophectoderm biopsy, Embryo biopsy,
Online Access:	http://hdl.handle.net/20.500.12792/1475 http://hdl.handle.net/10261/293481
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