Effects of serum albumin binding molecules on embryo culture and analytical system development for the molecules

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To evaluate small molecules non-specifically or specifically bound to the human serum albumin (HSA) including commercially available samples and human serum, we evaluated the small molecules before and after purification of HSA from human serum. The signals derived from bilirubin and cholesterol were detected. We found that a purification method using ethanol was necessary in order to remove bilirubin. 2-cell mouse embryo developed to blastocyst when purified HSA was added to the medium. Furthermore, when the mouse embryo is cultured in culture medium supplemented with fluorescence labeled bovine serum albumin (BSA), fluorescence of the BSA was observed in mouse blastocyst. Based on these results, we considered that fertilized oocyte could uptake the HSA in mouse embryonic development. Even if organic molecules bound to HSA, mouse embryonic development was observed. Because removal of binding molecules such as bilirubin and cholesterol is difficult without addition of salt and ethanol that affect the human embryo culture, we concluded that the HSA purified from human serum was not able to use for the embryo culture.