title: Improved Murine Blastocyst Quality and Development in a Single Culture Medium Compared to Sequential Culture Media.
abstract: OBJECTIVE
  Validate single versus sequential culture media for murine embryo development.
  
  
  METHODS
  Prospective laboratory experiment.
  
  
  METHODS
  Assisted Reproduction Laboratory.
  
  
  METHODS
  Murine embryos.
  
  
  METHODS
  Thawed murine zygotes cultured for 3 or 5 days (d3 or d5) in single or sequential embryo culture media developed for human in vitro fertilization.
  
  
  METHODS
  On d3, zygotes developing to the 8 cell (8C) stage or greater were quantified using 4',6-diamidino-2-phenylindole (DAPI), and quality was assessed by morphological analysis. On d5, the number of embryos reaching the blastocyst stage was counted. DAPI was used to quantify total nuclei and inner cell mass nuclei. Localization of ubiquitin C-terminal hydrolase L1 (UCHL1) and ubiquitin C-terminal hydrolase L3 (UCHL3) was reference points for evaluating cell quality.
  
  
  RESULTS
  Comparing outcomes in single versus to sequential media, the odds of embryos developing to the 8C stage on d3 were 2.34 time greater (P = .06). On d5, more embryos reached the blastocyst stage (P = <.0001), hatched, and had significantly more trophoblast cells (P = .005) contributing to the increased total cell number. Also at d5, localization of distinct cytoplasmic UCHL1 and nuclear UCHL3 was found in high-quality hatching blastocysts. Localization of UCHL1 and UCHL3 was diffuse and inappropriately dispersed throughout the cytoplasm in low-quality nonhatching blastocysts.
  
  
  CONCLUSIONS
  Single medium yields greater cell numbers, an increased growth rate, and more hatching of murine embryos. Cytoplasmic UCHL1 and nuclear UHCL3 localization patterns were indicative of embryo quality. Our conclusions are limited to murine embryos but one might speculate that single medium may also be more beneficial for human embryo culture. Human embryo studies are needed.
authors.affiliation: Division of Reproductive and Perinatal Research, Department of Obstetrics, Gynecology and Women's Health, School of Medicine, Columbia, MO, USA Division of Animal Sciences, College of Agriculture, Food and Natural Resources, Columbia, MO, USA.
authors.forename: Justin M
authors.initials: JM
authors.lastname: Hennings
authors.affiliation: Division of Reproductive and Perinatal Research, Department of Obstetrics, Gynecology and Women's Health, School of Medicine, Columbia, MO, USA.
authors.forename: Randall L
authors.initials: RL
authors.lastname: Zimmer
authors.affiliation: Division of Reproductive and Perinatal Research, Department of Obstetrics, Gynecology and Women's Health, School of Medicine, Columbia, MO, USA.
authors.forename: Henda
authors.initials: H
authors.lastname: Nabli
authors.affiliation: Department of Health Management and Informatics, School of Medicine, The University of Missouri, Columbia, MO, USA Department of Statistics, College of Arts and Sciences, The University of Missouri, Columbia, MO, USA Biostatistics and Research Design, Galena Hall, Columbia, MO, USA.
authors.forename: J Wade
authors.initials: JW
authors.lastname: Davis
authors.affiliation: Division of Reproductive and Perinatal Research, Department of Obstetrics, Gynecology and Women's Health, School of Medicine, Columbia, MO, USA Division of Animal Sciences, College of Agriculture, Food and Natural Resources, Columbia, MO, USA.
authors.forename: Peter
authors.initials: P
authors.lastname: Sutovsky
authors.affiliation: Division of Animal Sciences, College of Agriculture, Food and Natural Resources, Columbia, MO, USA.
authors.forename: Miriam
authors.initials: M
authors.lastname: Sutovsky
authors.affiliation: Division of Reproductive and Perinatal Research, Department of Obstetrics, Gynecology and Women's Health, School of Medicine, Columbia, MO, USA Division of Animal Sciences, College of Agriculture, Food and Natural Resources, Columbia, MO, USA timmsk@health.missouri.edu.
authors.forename: Kathy L
authors.initials: KL
authors.lastname: Sharpe-Timms
mesh_terms: D000818:Animals; D001755:Blastocyst / Q000187:drug effects* / Q000502:physiology*; D003470:Culture Media / Q000494:pharmacology*; D046149:Embryo Culture Techniques / Q000379:methods* / Q000592:standards; D047108:Embryonic Development / Q000187:drug effects* / Q000502:physiology*; D005260:Female; D006801:Humans; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL
chemical_list: D003470:Culture Media
keywords: UCHL1; UCHL3; culture; embryo; media