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Association of in vitro fertilization with global and IGF2/H19 methylation variation in newborn twins

Published online by Cambridge University Press:  10 April 2015

Y. J. Loke*
Early Life Epigenetics Group, Murdoch Childrens Research Institute (MCRI), Royal Children’s Hospital, Parkville, VIC, Australia
J. C. Galati
Clinical Epidemiology and Biostatistics Unit, MCRI, Royal Children’s Hospital, Parkville, VIC, Australia
R. Saffery
Cancer, Disease and Developmental Epigenetics Group, MCRI, Royal Children’s Hospital, Parkville, VIC, Australia Department of Paediatrics, University of Melbourne, Parkville, VIC, Australia
J. M. Craig
Early Life Epigenetics Group, Murdoch Childrens Research Institute (MCRI), Royal Children’s Hospital, Parkville, VIC, Australia Department of Paediatrics, University of Melbourne, Parkville, VIC, Australia
*Address for correspondence: Y. J. Loke, Early Life Epigenetics, Murdoch Childrens Research Institute (MCRI), Royal Children’s Hospital, Flemington Rd, Parkville, VIC 3052, Australia. (Email


In vitro fertilization (IVF) and its subset intracytoplasmic sperm injection (ICSI), are widely used medical treatments for conception. There has been controversy over whether IVF is associated with adverse short- and long-term health outcomes of offspring. As with other prenatal factors, epigenetic change is thought to be a molecular mediator of any in utero programming effects. Most studies focused on DNA methylation at gene-specific and genomic level, with only a few on associations between DNA methylation and IVF. Using buccal epithelium from 208 twin pairs from the Peri/Postnatal Epigenetic Twin Study (PETS), we investigated associations between IVF and DNA methylation on a global level, using the proxies of Alu and LINE-1 interspersed repeats in addition to two locus-specific regulatory regions within IGF2/H19, controlling for 13 potentially confounding factors. Using multiple correction testing, we found strong evidence that IVF-conceived twins have lower DNA methylation in Alu, and weak evidence of lower methylation in one of the two IGF2/H19 regulatory regions and LINE-1, compared with naturally conceived twins. Weak evidence of a relationship between ICSI and DNA methylation within IGF2/H19 regulatory region was found, suggesting that one or more of the processes associated with IVF/ICSI may contribute to these methylation differences. Lower within- and between-pair DNA methylation variation was also found in IVF-conceived twins for LINE-1, Alu and one IGF2/H19 regulatory region. Although larger sample sizes are needed, our results provide additional insight to the possible influence of IVF and ICSI on DNA methylation. To our knowledge, this is the largest study to date investigating the association of IVF and DNA methylation.

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© Cambridge University Press and the International Society for Developmental Origins of Health and Disease 2015 

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