title: In vitro maturation of canine oocytes co-cultured with bovine and canine granulosa cell monolayers.
abstract: The present study investigated the effects of bovine granulosa cell monolayers (BGML) and canine granulosa cell monolayers (CGML) on nuclear maturation of canine oocytes with and without cumulus cells. Cumulus-oocyte complexes (COCs) or cumulus-free oocytes were cultured in Dulbecco's Modified Eagle's Medium (DMEM, control group), DMEM with BGML (BGML group), or DMEM with CGML (CGML group) for 72 h at 38.5 °C in 5% CO(2), 5% O(2,) and 90% N(2). All media were supplemented with 10% of FCS, 50 ng/mL of EGF, 2 μg/mL of estradiol-17β, 0.1 IU/mL of hCG, 0.1 IU/mL of FSH, 0.25 mM of pyruvic acid, 100 μM of β-mercaptoethanol, 100 IU/mL of penicillin, and 100 μg/mL of streptomycin. In cumulus-enclosed oocytes retrieved from ovaries at estrus and/or diestrus, the highest percentage of M-II oocytes (P < 0.05) was present in the BGML group (27.0%) compared with the CGML group (7.9%) and the control group (3.5%). In cumulus-free oocytes collected from ovaries at estrus and/or diestrus, the proportions of M-II oocytes co-cultured with the CGML were low (3.0%) and similar (P > 0.05) to proportions achieved with control (3.0%). However, the presence of BGML improved (P < 0.05) the ability of denuded oocytes to develop into M-II (10.2%). The BGML group had the highest overall meiotic resumption (P < 0.05), and least oocyte degeneration (P < 0.05) among experimental groups. In conclusion, BGML had a positive impact on the in vitro maturation system, as well as meiotic resumption of canine oocytes.
authors: Research Unit for Functional Genomics, National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Inada, Obihiro, Japan.
authors: Mohammed Ali
authors: MA
authors: Abdel-Ghani
authors: Takashi
authors: T
authors: Shimizu
authors: Tomoyoshi
authors: T
authors: Asano
authors: Hiroshi
authors: H
authors: Suzuki
mesh_terms: D000818:Animals; D002417:Cattle; D002467:Cell Nucleus / Q000502:physiology; D002478:Cells, Cultured; D018920:Coculture Techniques; D054885:Cumulus Cells / Q000502:physiology; D004285:Dogs; D005260:Female; D006107:Granulosa Cells / Q000502:physiology*; D008540:Meiosis; D009865:Oocytes / Q000254:growth & development* / Q000648:ultrastructure