Effect of bone morphogenetic protein-4 on in vitro growth, steroidogenesis and subsequent developmental competence of the oocyte-granulosa cell complex derived from bovine early antral follicles.

Abstract

BackgroundBone morphogenetic proteins (BMPs) play important regulatory roles during folliculogenesis. Theca-derived BMP-4 may be beneficial to in vitro growth culture of early antral follicle-derived oocyte-granulosa cell complexes (OGCs), which is lacking in theca-derived products.MethodsBMP-4 (0 [control], 10 and 50 ng/mL) was added to growth culture medium. Growth, steroidogenesis and the subsequent developmental competence of OGCs derived from bovine early antral follicles (0.5-1 mm) were examined.ResultsAt 4, 8 and 12 days of growth culture, progesterone production by granulosa cells was suppressed by the addition of BMP-4 compared to the control (P < 0.05). At 12 days, both the OGC survivability and granulosa cell number in the 50 ng/mL BMP-4 treated group were lower than those of control (48.2 % vs. 67.8 %; 4.96 × 104 vs. 8.5 × 104 cells; P < 0.05, respectively), while no difference was found between 10 ng/mL and the control. The mean diameters of granulosa cell in the BMP-4 treated groups were smaller than that of the control (P < 0.05). However, the granulosa cell viability, oocyte diameter, oocyte nuclear maturation rate and normal fertilization rate were similar in all of the experimental groups, regardless of the amount of BMP-4 addition (P ˃ 0.05). BMP-4 treated in vitro-grown oocytes showed lower blastocyst rates than untreated ones (P < 0.05).ConclusionsBMP-4 addition during in vitro growth culture suppressed progesterone production and decreased the diameter of granulosa cells, suggesting its effect on steroidogenesis; importantly, it did not affect oocyte growth, nuclear maturation and fertilization. However, BMP-4 impaired subsequent embryonic development, and in higher concentration (50 ng/mL) even compromised OGC viability by suppressing proliferation of granulosa cells.