title: Development of a mouse model for studying the effect of embryo culture on embryonic stem cell derivation.
abstract: For most of the derived human embryonic stem cell (ESC) lines thus far, the majority of human embryos used have been frozen in liquid nitrogen at or prior to the compacting stage for up to 10 years before human ESC derivation. As such they were grown in media that were relatively simple in their formulation compared with those used today. Here we report that culture of mouse embryos in media similar to these produces blastocysts in which both the inner cell mass cell number and the number of ESC progenitor cells (epiblast cells) in the inner cell mass are reduced compared with blastocysts cultured in a purpose-designed sequential (G1/G2) system commonly used today. Embryos cultured in a simple medium were less likely to attach and generate outgrowths. Further, these outgrowths had increased metabolic activity, which has been linked to differentiation, and altered gene expression. Culture of embryos in a simple medium to the compacting stage followed by culture in G2 to the blastocyst stage reduced some of these effects. However, none were improved to the level seen for culture in G1/G2. These results highlight the influence of embryo culture on embryo quality and pluripotency, which is a key factor in determining ESC isolation efficiencies.
authors.affiliation: School of Paediatrics and Reproductive Health, University of Adelaide, Adelaide, South Australia.
authors.forename: Jared M
authors.initials: JM
authors.lastname: Campbell
authors.forename: Megan
authors.initials: M
authors.lastname: Mitchell
authors.forename: Mark B
authors.initials: MB
authors.lastname: Nottle
authors.forename: Michelle
authors.initials: M
authors.lastname: Lane
mesh_terms: D000818:Animals; D053624:Blastocyst Inner Cell Mass / Q000166:cytology* / Q000378:metabolism; D003470:Culture Media; D004265:DNA Helicases / Q000235:genetics / Q000378:metabolism; D046149:Embryo Culture Techniques; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000502:physiology; D020869:Gene Expression Profiling; D018398:Homeodomain Proteins / Q000378:metabolism; D053078:Membrane Potential, Mitochondrial; D051379:Mice; D008810:Mice, Inbred C57BL; D008808:Mice, Inbred CBA; D000071317:Nanog Homeobox Protein; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism; D051266:Nuclear Respiratory Factor 1 / Q000235:genetics / Q000378:metabolism; D056564:Sirtuin 1 / Q000235:genetics / Q000378:metabolism; D014327:Trophoblasts / Q000166:cytology / Q000378:metabolism; D000075924:X-linked Nuclear Protein
chemical_list: D003470:Culture Media; D018398:Homeodomain Proteins; D000071317:Nanog Homeobox Protein; C475540:Nanog protein, mouse; C495634:Nrf1 protein, mouse; D009687:Nuclear Proteins; D051266:Nuclear Respiratory Factor 1; C470545:Sirt1 protein, mouse; D056564:Sirtuin 1; D004265:DNA Helicases; C490462:Atrx protein, mouse; D000075924:X-linked Nuclear Protein; C491758:lymphoid specific helicase, mouse