Effects of cilostamide and forskolin on the meiotic resumption and embryonic development of immature human oocytes.
BACKGROUND
In an attempt to allow for acquisition of oocyte cytoplasmic maturation, PDE3 specific inhibitor, cilostamide and adenylate cyclase activator, forskolin were used to extend pre-maturation culture of immature human oocytes.


METHODS
Cumulus-oocyte complexes retrieved from unstimulated ovaries were continuously cultured under 20 microM cilostamide or 50 microM forskolin, alone or in combination for 6, 12, 24 or 48 h, respectively. Levels of intercellular gap junction communication (GJC) and maturational status were examined at these designated time points. Metaphase II oocytes obtained following 54 h biphasic culture (with meiotic inhibitors from 0 to 24 h, no meiotic inhibitors from 24 to 54 h) were subject to intracytoplasmic sperm injection and embryos were cultured for five more days.


RESULTS
Both cilostamide and forskolin delayed spontaneous meiotic progression after continuous culture with immature human oocytes. Combined treatment of cilostamide and forskolin significantly lowered the rates of germinal vesicle breakdown (GVBD) at 6, 12, 24 or 48 h after meiotic inhibitory culture, when compared with the control (all P < 0.05). A delay of 6 h for the loss of GJC was also observed under the combined treatment of cilostamide and forskolin. The fertilization rate was significantly higher under the combined treatment of cilostamide and forskolin than that of the control. Although the rates of oocyte maturation and embryo cleavage were similar among groups, there was a slight but non-significant increase in blastocyst formation rate with the treatment of cilostamide and forskolin.


CONCLUSIONS
Combined treatment of cilostamide and forskolin positively influences oocyte developmental competence by exhibiting a synergistic effect on the prevention of GJC loss and resumption of meiosis.
Shu
Yi-min
YM
Center for Reproductive Medicine, Department of Obstetrics and Gynecology, the First Affiliated Hospital, Sun Yat-sen University, Guangzhou 510080, People's Republic of China. shuyimin@hotmail.com
Zeng
Hai-tao
HT
Ren
Zi
Z
Zhuang
Guang-lun
GL
Liang
Xiao-Yan
XY
Shen
Hong-wei
HW
Yao
Shu-zhong
SZ
Ke
Pei-qi
PQ
Wang
Ning-ning
NN
D000262:Adenylyl Cyclases / Q000378:metabolism; D000328:Adult; D002478:Cells, Cultured; D005576:Colforsin / Q000494:pharmacology*; D004357:Drug Synergism; D047108:Embryonic Development / Q000187:drug effects*; D005260:Female; D017629:Gap Junctions / Q000187:drug effects / Q000502:physiology; D006801:Humans; D008540:Meiosis / Q000187:drug effects*; D009865:Oocytes / Q000166:cytology* / Q000187:drug effects*; D010726:Phosphodiesterase Inhibitors / Q000494:pharmacology; D015363:Quinolones / Q000494:pharmacology*
D010726:Phosphodiesterase Inhibitors; D015363:Quinolones; D005576:Colforsin; C021294:cilostamide; D000262:Adenylyl Cyclases