In vitro maturation of bitch oocytes from advanced preantral follicles in synthetic oviduct fluid medium: serum is not essential

Abstract

The ability of oocytes from preantral follicles to mature in vitro was assessed using a synthetic oviduct fluid (SOF) medium. Advanced preantral follicles (approximately 210μm diameter) were isolated from the ovaries of domestic bitches and assigned to one of four treatment groups: (1) SOF (n=230); (2) SOF+3mg/ml bovine serum albumin (+BSA, n=220); (3) SOF+20% fetal bovine serum (+FBS, n=227); or (4) SOF+3mg/ml BSA+20% FBS (+BSA+FBS, n=232), then cultured for up to 72h. A group of control follicles was not cultured (n=103). The percentages of oocytes reaching metaphase I to metaphase II stages (MI to MII) did not differ between treatments at each culture period. Within treatments, the percentages of oocytes at MI to MII stages did not differ with duration of culture. However, when compared to the control group (0.97%) the percentages of oocytes at MI to MII increased (P<0.05) in the SOF group after 48h (10.0%) and 72h (12.2%) of culture. In the +BSA (10.1%) and +FBS (9.7%) groups, the percentages of oocytes at MI to MII increased (P<0.05) above control values only after 72h of culture. The percentage of oocytes at MI to MII did not significantly increase in the +BSA+FBS group (3.9, 6.6 and 7.6% at 24, 48 and 72h of culture, respectively) compared to the control group. These results indicate that under the described conditions supplementation of culture medium with BSA or FBS is not essential, and the simple medium SOF can support nuclear maturation of a small proportion of bitch oocytes in vitro.