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Human serum albumin (HSA) is usually added to human in vitro fertilization culture medium to support embryo development, but carries some risk of contamination with various other compounds. Recombinant human albumin (rHA) has been shown to be a chemically defined protein. We evaluated the effect of rHA on mouse embryo development.B6D2F1 pronuclear oocytes were cultured in protein-free potassium simplex optimized medium with non-essential and essential amino acids supplemented with rHA, HSA (1 mg/mL) or polyvinyl alcohol (PVA; 0.1 mg/mL) for 96 h. The incidence of apoptosis and the generation of nitric oxide (NO) in blastocysts and fetal development after embryo transfer were examined.Blastocyst development was equal in the three supplements. The incidence of apoptosis and the generation of NO in blastocysts developed in rHA was significantly (P < 0.05) lower than in HSA or PVA. After transfer of blastocysts developed in rHA, the percentage of fetal development was significantly (P < 0.05) higher than in HSA or PVA (75.8 ± 2.2% vs 46.8 ± 7.5% or 42.4 ± 3.9%, respectively).Supplementing culture medium with rHA resulted in decreased apoptosis and increased fetal development after embryo transfer. The results show that the presence of rHA in the culture medium is beneficial for producing high-quality blastocysts.