Growing porcine oocyte-granulosa cell complexes acquired meiotic competence during in vitro culture.
The aim of this study was to establish a culture system to improve the meiotic competence of porcine oocyte-granulosa cell complexes (OGCs) obtained from preantral or early antral follicles. Porcine OGCs were recovered from follicles with diameters of 230-300 (preantral follicles), 300-500, and 500-700 mum (early antral follicles) using scalpels. The OGCs were cultured for 2 weeks in culture medium. We examined the effects of the sizes of the follicles from which OGCs were recovered, the concentrations of polyvinylpyrrolidone (PVP, 0-8%) in the culture medium, and 2 types of culture dish (Falcon 3002 vs 1007) on formation of the antrum of OGCs. After culture, the oocytes were matured for 44 h to assess their meiotic competence. OGCs recovered from small follicles (230-500 microm) required longer (P<0.05) than larger follicles to form the antrum structure. The percentage of OGCs forming the antrum structure that were cultured in 2% PVP (31%) was higher (P<0.05) than for those cultured in other PVP concentrations (0-11%). The percentages of antrum-structure formation for OGCs cultured on Falcon 3002 (83% for 2% PVP and 60% for 4% PVP) were higher (P<0.05) than those cultured on Falcon 1007 (47% for 2% PVP and 9% for 4% PVP). Furthermore, all of the intact oocytes that were obtained from culture of OGCs and that formed an antrum were in the GV stage (n=28). When these immature oocytes were cultured for 44 h, the percentage of oocytes that reached the metaphase II stage (25%, n=68) was higher (P<0.0001) than that of oocytes matured without culture (0.7%, n=137). The results of the present study show that porcine OGCs obtained from preantral or early antral follicles acquire meiotic competence in vitro.
Hashimoto
Shu
S
IVF Namba Clinic-Osaka, Japan. hashimoto@ivfnamba.com
Ohsumi
Kanako
K
Tsuji
Yoko
Y
Harauma
Naoko
N
Miyata
Yuko
Y
Fukuda
Aisaku
A
Hosoi
Yoshihiko
Y
Iritani
Akira
A
Morimoto
Yoshiharu
Y
D000818:Animals; D018929:Cell Culture Techniques; D048429:Cell Size; D002478:Cells, Cultured; D003470:Culture Media / Q000737:chemistry / Q000494:pharmacology; D005260:Female; D006107:Granulosa Cells / Q000166:cytology* / Q000502:physiology*; D008540:Meiosis; D009865:Oocytes / Q000166:cytology* / Q000502:physiology*; D011205:Povidone / Q000494:pharmacology; D034421:Sus scrofa
D003470:Culture Media; D011205:Povidone