title: Side-by-side comparison of five commercial media systems in a mouse model: suboptimal in vitro culture interferes with imprint maintenance.
abstract: Assisted reproductive technologies (ARTs) are becoming increasingly prevalent and are generally considered to be safe medical procedures. However, evidence indicates that embryo culture may adversely affect the developmental potential and overall health of the embryo. One of the least studied but most important areas in this regard is the effects of embryo culture on epigenetic phenomena, and on genomic imprinting in particular, because assisted reproduction has been linked to development of the human imprinting disorders Angelman and Beckwith-Wiedemann syndromes. In this study, we performed side-by-side comparisons of five commercial embryo culture systems (KSOMaa, Global, Human Tubal Fluid, Preimplantation 1/Multiblast, and G1v5PLUS/G2v5PLUS) in relation to a best-case (in vivo-derived embryos) and a worst-case (Whitten culture) scenario. Imprinted DNA methylation and expression were examined at three well-studied loci, H19, Peg3, and Snrpn, in mouse embryos cultured from the 2-cell to the blastocyst stage. We show that embryo culture in all commercial media systems resulted in imprinted methylation loss compared to in vivo-derived embryos, although some media systems were able to maintain imprinted methylation levels more similar to those of in vivo-derived embryos in comparison to embryos cultured in Whitten medium. However, all media systems exhibited loss of imprinted H19 expression comparable to that using Whitten medium. Combined treatment of superovulation and embryo culture resulted in increased perturbation of genomic imprinting, above that from culture alone, indicating that multiple ART procedures further disrupt genomic imprinting. These results suggest that time in culture and number of ART procedures should be minimized to ensure fidelity of genomic imprinting during preimplantation development.
authors: Department of Obstetrics & Gynecology, University of Western Ontario, Schulich School of Medicine and Dentistry, London, Ontario, Canada.
authors: B A
authors: BA
authors: Market-Velker
authors: A D
authors: AD
authors: Fernandes
authors: M R W
authors: MR
authors: Mann
mesh_terms: D000818:Animals; D001755:Blastocyst / Q000378:metabolism*; D003470:Culture Media / Q000737:chemistry*; D019175:DNA Methylation; D004477:Ectogenesis; D046149:Embryo Culture Techniques / Q000379:methods*; D005260:Female; D005306:Fertilization; D005307:Fertilization in Vitro / Q000009:adverse effects; D018392:Genomic Imprinting; D051741:Kruppel-Like Transcription Factors / Q000235:genetics / Q000378:metabolism; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D062085:RNA, Long Noncoding; D022661:RNA, Untranslated / Q000235:genetics / Q000378:metabolism; D027724:Reproductive Techniques, Assisted / Q000009:adverse effects; D013223:Statistics as Topic; D013480:Superovulation; D055517:snRNP Core Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D003470:Culture Media; C094335:H19 long non-coding RNA; D051741:Kruppel-Like Transcription Factors; C097901:Peg3 protein, mouse; D062085:RNA, Long Noncoding; D022661:RNA, Untranslated; D055517:snRNP Core Proteins