In vitro maturation of bitch oocytes from advanced preantral follicles in synthetic oviduct fluid medium: serum is not essential.
The ability of oocytes from preantral follicles to mature in vitro was assessed using a synthetic oviduct fluid (SOF) medium. Advanced preantral follicles (approximately 210 microm diameter) were isolated from the ovaries of domestic bitches and assigned to one of four treatment groups: (1) SOF (n = 230); (2) SOF + 3 mg/ml bovine serum albumin (+BSA, n = 220); (3) SOF + 20% fetal bovine serum (+FBS, n = 227); or (4) SOF + 3 mg/ml BSA + 20% FBS (+BSA+FBS, n = 232), then cultured for up to 72 h. 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 48 h (10.0%) and 72 h (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 72 h 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 72 h 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.
Bolamba
Digbo
D
Reproductive Physiology Division, Center for Reproduction of Endangered Species, Zoological Society of San Diego, PO Box 120551, San Diego, CA 92112-0551, USA.
Russ
Kara D
KD
Olson
Mary A
MA
Sandler
Jody L
JL
Durrant
Barbara S
BS
D000818:Animals; D001826:Body Fluids; D002478:Cells, Cultured; D003470:Culture Media; D016895:Culture Media, Serum-Free; D004285:Dogs; D005187:Fallopian Tubes / Q000378:metabolism*; D005260:Female; D008677:Metaphase; D009865:Oocytes / Q000502:physiology* / Q000648:ultrastructure; D006080:Ovarian Follicle / Q000166:cytology*; D012710:Serum Albumin, Bovine
D003470:Culture Media; D016895:Culture Media, Serum-Free; D012710:Serum Albumin, Bovine