Characterization of secreted proteins of 2-cell mouse embryos cultured in vitro to the blastocyst stage with and without protein supplementation

J Assist Reprod Genet. 2014 Jun;31(6):757-65. doi: 10.1007/s10815-014-0207-2.


Purpose: To identify the secreted proteins of murine embryos grown in vitro.

Methods: Two-cell mouse embryos (n=432) were randomly allocated to culture to the blastocyst stage in protein-free and in protein-supplemented (3 % BSA) media. Proteins were separated by SDS-PAGE; bands were visualized by coomassie staining, followed by in-gel trypsin digestion and liquid chromatography-tandem mass spectrometry. RT-PCR and confocal microscopy were used to confirm gene/protein expression in blastocysts.

Results: Of all individually identified proteins, 34 and 23 were found in embryos cultured without and with BSA, respectively, and 20 were common. Identified proteins having an N-terminal secretory sequence or transmembrane domains located on the extracellular backbone were postulated as secreted proteins. Gene and protein expression for two selected molecules were confirmed. Functional analysis revealed over-represented processes related to lipid metabolism, cyclase activity, and cell adhesion/membrane functions.

Conclusions: This study provided evidence to further characterize secreted proteins by mouse embryos grown from the 2-cell to the blastocyst stage in vitro. Because of homology between murine and human, these results may provide information to be translated to the clinical setting.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Blastocyst / cytology*
  • Culture Media / chemistry
  • Embryo, Mammalian / drug effects
  • Embryo, Mammalian / metabolism*
  • Embryonic Development / drug effects
  • Gene Expression Regulation, Developmental / drug effects
  • Humans
  • Mice
  • Protein Biosynthesis / genetics*
  • Proteins / administration & dosage*
  • Proteins / chemistry


  • Culture Media
  • Proteins