title: Variability in protein quality used for embryo culture: embryotoxicity of the stabilizer octanoic acid.
abstract: OBJECTIVE
  To screen human serum albumin (HSA) preparations for toxicity and investigate causes of variation.
  
  
  METHODS
  Experimental laboratory study.
  
  
  METHODS
  University-based laboratory.
  
  
  METHODS
  FVB and CF1 mice crossed to create embryos used in experiments.
  
  
  METHODS
  Mouse embryo assay performed with 5% or 15% HSA (100 mg/mL albumin) from three samples from three separate manufacturers (A, B, C).
  
  
  METHODS
  Blastocyst rates calculated at 96 hours of culture (experiments repeated in triplicate).
  
  
  RESULTS
  The HSA preparations were desalted to remove stabilizers added during HSA processing, then mass spectrometry was used to determine the relative variation in stabilizer concentrations; the effect of the stabilizer octanoic acid on embryo development was tested. At 5% HSA, all samples had blastocyst rates ≥ 70%; at 15% HSA, the blastocyst rates for samples B and C were <50%. Desalting did not affect sample B but did improve the blastocyst rates of sample C. Mass spectrometry revealed high levels of octanoic acid in sample C compared with sample A. The addition of octanoic acid to sample A produced toxicity similar to sample C.
  
  
  CONCLUSIONS
  The stabilizer octanoic acid varies by lot and inhibits embryo development. Because octanoic acid is known to cause disruptions in mitochondrial bioenergetics, reduce intracellular pH, and induce oxidative damage in peripheral tissues, its use in embryo culture should be monitored and limited.
authors.affiliation: Department of Obstetrics and Gynecology, Division of Reproductive Endocrinology and Infertility, Mayo Clinic, Rochester, Minnesota 55905, USA.
authors.forename: Phoebe H
authors.initials: PH
authors.lastname: Leonard
authors.forename: M Cristine
authors.initials: MC
authors.lastname: Charlesworth
authors.forename: Linda
authors.initials: L
authors.lastname: Benson
authors.forename: David L
authors.initials: DL
authors.lastname: Walker
authors.forename: Jolene R
authors.initials: JR
authors.lastname: Fredrickson
authors.forename: Dean E
authors.initials: DE
authors.lastname: Morbeck
mesh_terms: D000818:Animals; D002210:Caprylates / Q000633:toxicity*; D003470:Culture Media / Q000737:chemistry / Q000592:standards* / Q000633:toxicity; D004305:Dose-Response Relationship, Drug; D004355:Drug Stability; D046149:Embryo Culture Techniques / Q000379:methods / Q000592:standards* / Q000706:statistics & numerical data; D004622:Embryo, Mammalian / Q000166:cytology / Q000187:drug effects*; D005079:Excipients / Q000633:toxicity*; D005260:Female; D006801:Humans; D051379:Mice; D011786:Quality Control; D012492:Salts / Q000737:chemistry; D012709:Serum Albumin / Q000737:chemistry / Q000494:pharmacology / Q000633:toxicity
chemical_list: D002210:Caprylates; D003470:Culture Media; D005079:Excipients; D012492:Salts; D012709:Serum Albumin; C031492:octanoic acid
keywords: Albumin; embryo culture; mouse embryo assay; quality control; toxicity