A simple method for selection of cumulus-oocyte complexes from bovine ovaries by sedimentation with percoll.
Selection of bovine cumulus-oocyte complexes (COCs) for in vitro embryo production (IVP) is generally based on the morphological characteristics of the cumulus cells surrounding the oocytes and the ooplasm under microscopic observation. The purpose of this study was to examine a simple method for selection of COCs by sedimentation with Percoll solutions. COCs were aspirated from ovaries derived from a local slaughterhouse, and the COCs were classified by the morphology of their cumulus cell layers, as follows: Class A, compact and thick; Class B, compact but thin; Class C, partially denuded and thin; and Class D, denuded. Percoll solutions were prepared by diluting Percoll to 7.5, 10, 12.5, 15, 17.5, 20, 22.5, 25, 27.5 and 30% solutions, respectively. COCs were placed on the surface of the Percoll solution for 3 min, and the precipitated COCs were transferred to stepwise high density solution. The percentage of Percoll solution just before buoyancy was considered to the specific sedimentary value of the COC and oocyte. The mean sedimentary value of Class A COCs was higher than those of the other classes (P<0.01). The mean sedimentary values of denuded oocytes from Classes A and B were higher than those from Classes C and D (P<0.01). Our results show that sedimentation of COCs and denuded oocytes was generally related to the morphological quality of the COCs, although the sedimentary values ranged widely for one class of COCs and oocytes. The Percoll method can be used for simple selection of COCs.
Yotsushima
Kenji
K
Livestock Experiment Station, Toyama Prefectural Agricultural Research Center, Toyama, Japan. kenji.yotsushima@pref.toyama.lg.jp
Shimizu
Masayo
M
Kon
Hiroaki
H
Izaike
Yoshiaki
Y
D000818:Animals; D002417:Cattle; D002469:Cell Separation / Q000379:methods*; D005260:Female; D005307:Fertilization in Vitro / Q000662:veterinary*; D009865:Oocytes / Q000166:cytology*; D010053:Ovary / Q000166:cytology; D011205:Povidone; D012822:Silicon Dioxide
C016039:Percoll; D012822:Silicon Dioxide; D011205:Povidone