Spindle and chromosome configurations of human oocytes matured in vitro in two different culture media

Abstract 

In-vitro maturation can have deleterious effects on spindle formation and proper chromosome alignment in human oocytes and can be profoundly affected by culture conditions. This study compared the spindle presence and location with the maturation rate of germinal vesicle (GV) oocytes cultured in two different media: G1.2 and G1.2 supplemented with follicle-stimulating hormone, human chorionic gonadotrophin and 17β-oestradiol. A total of 304 oocytes were retrieved from 101 women undergoing IVF treatment with intracytoplasmic sperm injection. Spindle presence was recorded using the Polscope. Spindle morphology was evaluated with immunocytological staining for α-tubulin and chromatin. Twenty-one in-vitro matured oocytes with the presence of spindle and ten of their corresponding polar bodies (PB) were also assessed for aneuploidy. A significantly increased maturation rate (69.7%) was observed after 24h in the supplemented culture media compared with the G1.2 media (56.6%; P<0.05). The proportions of metaphase II (MII) oocytes with spindle presence and abnormal spindle morphology were similar in the two culture media. Also, 76.9% of MII and 70% of PB had chromosomal abnormalities. In conclusion, supplementing culture media may increase the oocyte maturation rate in vitro, but does not necessarily indicate the presence of a birefringent spindle, or normal spindle and chromosomal alignment.

Keywords: culture medium, FISH, immunostaining, oocyte maturation, polscope, spindles