The objective of this study was to determine the effect of bovine follicular fluid proteins (bFF) and their differently charged groups as maturation media supplements for in vitro embryo development. bFF was obtained by aspiration from large healthy follicles (4–10 mm in diameter) and was precipitated by 30–50% (NH₄)₂SO₄. The precipitated protein was fractionated into basic and acidic fractions by ion-exchanger columns. In experiment 1, the oocytes were matured in TCM-199 with 1) FBS+hormones (control) and 2) 10% bFF. The oocyte maturation rate, the development to the blastocyst stage rate and blastocyst cell number were not significantly different between the groups. However, the INFα and IGF-2r expression levels in the 10% bFF were higher than in the control (P<0.05). In experiment 2, the specific charge proteins of bFF (basic and acidic) were also used as media supplements in the maturation medium. The basic fraction had higher oocyte maturation rate and blastocyst cell number when compared with addition of acidic fraction (P<0.05). The expression levels for almost all developmentally important genes in the basic fraction were greater than those in the acidic fraction, particularly INFα (P<0.05). Most of the protein in the basic fraction was associated with the immune response and mRNA processing. In conclusion, supplementation of 10% bFF alone in maturation medium can support oocyte maturation and embryo development. The basic fraction in bFF seemed to have effect on oocyte maturation rate and blastocyst cell number.