title: Effects of cumulus cells, different cryoprotectants, various maturation stages and preincubation before insemination on developmental capacity of frozen-thawed bovine oocytes.
abstract: To improve freezability of bovine follicular oocytes, it is necessary to minimize injury to the oocytes caused by freezing and the toxicity of cryoprotectants. The maturing ability of frozen-thawed follicular oocytes with or without cumulus complexes was tested. The proportion of frozen-thawed follicular oocytes reaching the metaphasc II (M II) stage after in vitro maturation of 24 h was significantly (P < 0.05) higher in cumulus oocyte complexes (COCs; 44%) than in denuded oocytes (30%). Oocytes were cultured for 0, 6, 12, 18 or 24 h then frozen-thawed with 1,2-propanediol (PROH) or dimethyl sulfoxide (DMSO), and cultured for 24, 18, 12, 6 or 0 h respectively. In PROH, 24:0 (67%) showed significantly (P < 0.05) higher maturation rate than 0:24 (38%), 6:18 (41%). In DMSO, 18:6 (72%) and 24:0 (61%) showed significantly (P < 0.05) higher maturation rate than 0:24 (30%), 6:18 (33%) and 12:12 (44%). In case of 18:6, DMSO (72%) showed significant (P < 0.05) higher maturation rate than PROH (52%), however in case of 0:24, 6:18, 12:12 and 24:0, there was no significant (P < 0.05) difference in the maturation rate between PROH and DMSO. The proportion of embryos developed to > or = 2 cell, > or = 8 cell, morula and blastocyst in 18:6 DMSO (35, 10, 3 and 0%) and 24:0 PROH (38, 12, 5 and 0%) was significantly (P < 0.05) lower than that of fresh oocytes (67, 38, 31 and 16%). There was no significant (P < 0.05) difference in the rate of embryos that developed to > or = 2 cells, > or = 8 cells, morulae and blastocysts between PROH and DMSO. When the frozen oocytes were grouped as rewarming culture (21:2 PROH) and control (24:0 PROH), there was no significant (P < 0.05) difference in the rate of embryos that developed to > or = 2 cells, > or = 8 cells, morulae and blastocysts between 24:0 PROH (42, 24, 11 and 1%) and 21:2 PROH (51, 29, 16 and 4%) but 21:2 PROH showed slightly higher developmental capacity than 24:0 PROH. Transferable blastocysts (4%) were obtained in 21:2 PROH when the frozen-thawed follicular oocytcs were fertilized and cultured for 8 to 9 d.
authors.affiliation: Department of Animal Science & Technology, College of Agriculture & Life Sciences, Seoul National University, Suweon 441-744, Korea.
authors.forename: K S
authors.initials: KS
authors.lastname: Im
authors.forename: J K
authors.initials: JK
authors.lastname: Kang
authors.forename: H S
authors.initials: HS
authors.lastname: Kim