Abstract
ObjectiveTo study whether methylated CpG-island (CGI) amplification coupled with microarray (MCAM) can be used to generate DNA (deoxyribonucleic acid) methylation profiles from single human blastocysts.DesignA pilot microarray study with methylated CpG-island amplification applied to human blastocyst genomic DNA and hybridized on CpG-island microarrays.SettingUniversity research laboratory.Patient(s)Five cryopreserved sibling 2-pronuclear zygotes that were surplus to requirements for clinical treatment by in vitro fertilization were donated with informed consent from a patient attending Bourn Hall Clinic, Cambridge, United Kingdom.Intervention(s)None.Main Outcome Measure(s)Successful generation of genome-wide DNA methylation profiles at CpG islands from individual human blastocysts, with common genomic regions of DNA methylation identified between embryos.Result(s)Between 472 and 734 CpG islands were methylated in each blastocyst, with 121 CpG islands being commonly methylated in all 5 blastocysts. A further 159 CGIs were commonly methylated in 4 of the 5 tested blastocysts. Methylation was observed at a number of CGIs within imprinted-gene, differentially methylated regions (DMRs), including placental and preimplantation-specific DMRs.Conclusion(s)The MCAM method is capable of providing comprehensive DNA methylation data in individual human blastocysts.
Highlights
We sought to identify whether genomewide DNA methylation analysis of CpG (C–phosphate–G) islands is feasible in single human blastocysts
Methylated CGI amplification coupled with microarray (MCAM) [12] is a method of DNA methylation analysis that has been used to assess CGI methylation in several human disease states [13,14,15]
We established that the MCAM method is suitable for CGI methylation analysis in individual human blastocysts
Summary
To study whether methylated CpG-island (CGI) amplification coupled with microarray (MCAM) can be used to generate. Defining the DNA methylome of the human preimplantation embryo may reveal which genomic regions may be susceptible to the effects of embryonic development in vitro, by identifying their requirement for a specific methylation state in early development. In this pilot study, we sought to identify whether genomewide DNA methylation analysis of CpG (C–phosphate–G) islands is feasible in single human blastocysts. We sought to identify whether genomewide DNA methylation analysis of CpG (C–phosphate–G) islands is feasible in single human blastocysts These islands (CGIs) are regions of the genome important in regulating gene expression in a manner dictated by their methylation status [10, 11]. Methylated CGI amplification coupled with microarray (MCAM) [12] is a method of DNA methylation analysis that has been used to assess CGI methylation in several human disease states [13,14,15]
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