Deoxyribonucleic acid methylation profiling of single human blastocysts by methylated CpG island amplification coupled with CpG island microarray

Capsule:
Methylated CpG-island amplification coupled with CpG-island microarray was used to assess DNA methylation in individual in vitro–derived human blastocysts, revealing 121 CpG islands that were hypermethylated in all 5 embryos.

Authors:
John Huntriss, Ph.D., Karen Hemmings, Ph.D., Praveen Baskaran, B.Sc., Lee Hazelwood, Ph.D., Kay Elder, Ph.D., Carl Virtanen, M.Sc., David Miller, Ph.D., Helen M. Picton, Ph.D.

Volume 103, Issue 6, Pages 1566-1571

Abstract:

Objective:
To 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.

Design:
A pilot microarray study with methylated CpG-island amplification applied to human blastocyst genomic DNA and hybridized on CpG-island microarrays.

Setting:
University 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.

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