title: Composition of protein supplements used for human embryo culture.
abstract: OBJECTIVE
  To determine the composition of commercially available protein supplements for embryo culture media and test if differences in protein supplement composition are biologically relevant in a murine model.
  
  
  METHODS
  Amino acid, organic acid, ion and metal content were determined for 6 protein supplements: recombinant human albumin (AlbIX), human serum albumin (HSA and Buminate), and three complex protein supplements (SSS, SPS, LGPS). To determine if differences in the composition of these supplements are biologically relevant, mouse one-cell embryos were collected and cultured for 120 hours in each protein supplement in Global media at 5 and 20 % oxygen in an EmbryoScope time-lapse incubator. The compositions of six protein supplements were analyzed for concentrations of 39 individual amino acids, organic acids, ions and elements. Blastocyst development and cell cycle timings were calculated at 96-hours of culture and the experiments were repeated in triplicate. Blastocyst gene expression was analyzed.
  
  
  RESULTS
  Recombinant albumin had the fewest undefined components , the lowest concentration of elements detected, and resulted in high blastocyst development in both 5 and 20 % oxygen. Buminate, LGPS and SPS had high levels of transition metals whereas SSS had high concentrations of amino acids. Pre-compaction mouse embryo development was delayed relative to embryos in AlbIX for all supplements and blastocyst formation was reduced in Buminate, SPS and SSS.
  
  
  CONCLUSIONS
  The composition of protein supplements are variable, consisting of previously undescribed components. High concentrations of pro-oxidant transition metals were most notable. Blastocyst development was protein dependent and showed an interaction with oxygen concentration and pro-oxidant supplements.
authors.affiliation: Department of Obstetrics and Gynecology, Mayo Clinic, Charlton 3A, Rochester, MN, 55905, USA, Morbeck.Dean@mayo.edu.
authors.forename: Dean E
authors.initials: DE
authors.lastname: Morbeck
authors.forename: Melissa
authors.initials: M
authors.lastname: Paczkowski
authors.forename: Jolene R
authors.initials: JR
authors.lastname: Fredrickson
authors.forename: Rebecca L
authors.initials: RL
authors.lastname: Krisher
authors.forename: Heather S
authors.initials: HS
authors.lastname: Hoff
authors.forename: Nikola A
authors.initials: NA
authors.lastname: Baumann
authors.forename: Thomas
authors.initials: T
authors.lastname: Moyer
authors.forename: Dietrich
authors.initials: D
authors.lastname: Matern
mesh_terms: D000596:Amino Acids / Q000737:chemistry / Q000302:isolation & purification; D000818:Animals; D001755:Blastocyst / Q000187:drug effects; D003470:Culture Media / Q000737:chemistry*; D046149:Embryo Culture Techniques / Q000379:methods*; D004622:Embryo, Mammalian; D047108:Embryonic Development / Q000187:drug effects*; D005307:Fertilization in Vitro; D006801:Humans; D007477:Ions / Q000737:chemistry / Q000302:isolation & purification; D008670:Metals / Q000737:chemistry / Q000302:isolation & purification; D051379:Mice; D017382:Reactive Oxygen Species / Q000378:metabolism; D012709:Serum Albumin / Q000737:chemistry / Q000494:pharmacology
chemical_list: D000596:Amino Acids; D003470:Culture Media; D007477:Ions; D008670:Metals; D017382:Reactive Oxygen Species; D012709:Serum Albumin