The Cosmic Evolution Survey (COSMOS) is a Hubble Space Telescope (HST) treasury project. The COSMOS aims to perform a 2 square degree imaging survey of an equatorial field in I(F814W) band, using the Advanced Camera for Surveys (ACS). Such a wide field survey, combined with ground-based photometric and spectroscopic data, is essential to understand the interplay between large scale structure, evolution and formation of galaxies and dark matter. In 2004, we have obtained high-quality, broad band images of the COSMOS field (B, V, r', i', and z') using Suprime-Cam on the Subaru Telescope, and we have started our new optical multi-band program, COSMOS-21 in 2005. Here, we present a brief summary of the current status of the COSMOS project together with contributions from the Subaru Telescope. Our future Subaru program, COSMOS-21, is also discussed briefly.

The objective of this study was to examine the effect of EGF on meiotic maturation and pronuclear (PN) formation of porcine oocytes. Prepubertal gilt cumulus-oocyte-complexes (COCs) aspirated from 2~6mm follicles of abbatoir ovaries were matured in TCM199 containing 0.1mg/ml cysteine, 0.5㎍/ml FSH and LH, and EGF (0, 5, 10, 20, 40 ng/ml) for 22 hr at 39℃ in a humidified atmosphere of 5% CO2 in air. They were then cultured for an additional 22hr without hormones. In Experiment 1, to examine the nuclear maturation at 44hr of culture, the expanded cumulus cells were removed by vortexing for 1 min in 3 mg/ml hyaluronidase. The oocytes were fixed in acetic acid: methanol (1:3, v/v) at least for 48 hr and stained with 1% orcein solution for 5 min. Nuclear status was classified as germinal vesicle (GV), germinal vesicle breakdown (GVBD), prophase-metaphase I (PI-MI), and PII-MII under microscope. In Experiment 2, to investigate PN formation, oocytes were fertilized with Percoll-treated freshly ejaculated sperm (1 x10 5 cells/ml) in mTBM with 0.3% BSA and 2mM caffeine for 5hr, and cultured in NCSU-23 medium with 0.4% BSA. At 6hr of culture, the embryos were fixed in 3.7% formaldehyde for 48hr and stained with 10ug/ml propidium iodide for 30 min. PN status was classified as no or one PN (unfertilized), 2 PN (normal fertilized) and ≥3 PN (polyspermy). Differences between groups were analyzed using one-way ANOVA after arc-sine transformation of the proportional data. The rate of oocytes that had reached to PII-MII were significantly (P<0.05) higher in all groups added EGF than that of non-treated group (67%), but it did not differ among the all added groups (86%, 85%, 79% and 81%, in 5, 10, 20 and 40 ng/ml EGF, respectively). No differences on the incidence of 2PN were observed in all treated groups (25%, 30%, 33%, 29% and 29%, in 0, 5, 10, 20 and 40 ng/ml EGF, respectively), however, in non-treated group, polyspermy tended to be increased (66% vs 58%, 54%, 52% and 55%, 0 vs. 5, 10, 20, 40 ng/ml EGF, respectively). These results suggest that EGF can be effectively used as an additive for enhancing oocyte maturation and reducing the incidence of polyspermy in pig.

The present study was performed to determine the ability of canine oocytes to achieve nuclear maturation according to oocyte diameter and different culture environments. All of the collected oocytes were classified by grade 1 to 3 and by their diameters such as <100㎛, <100㎛ to <110㎛, <110㎛ to <120㎛, >120㎛. Oocytes were cultured in culture medium supplemented with 10%FBS, 0.4%BSA,10% porcine follicular fluid (pFF), 10% canine serum (CS), or 10% canine estrus serum (CES). The mean number of oocytes recovered from estrus status ovaries was significantly higher than that of anestrus status ovaries (p<0.01). The maturation rate of grade 1 oocytes (>120㎛) was significantly higher than that of the other groups (p<0.05). Nuclear maturation to MI to MII in diameter of >110㎛ groups was significantly higher than that in <100㎛ group (p<0.05). The oocytes cultured in 10% FBS­supplemented group were significantly higher rate of GVBD compared to the other supplemented groups (p<0.05), and oocytes maturation to MI to MII in 10% FBS-, 0.4% BSA-, and 10% pFF-supplemented groups were significantly higher than those in 10% CS-supplemented group (p<0.05). Based on these results, the estrus status and the size of oocyte affect positively to improve nuclear maturation of canine immature oocytes in vitro. Among several protein sources, porcine follicular fluid was the most effective supplementation to culture medium to achieve higher in vitro maturation rate.

This study were examined whether plasminogen activators (PAs) are produced by porcine fresh or frozen-thawed cumulus-oocytes complexes (COCs) and cumulus cell free-oocytes. In fresh or frozen-thawed COCs and oocytes for 0 hour cultured, no activity of PAs was detected. However, at 24 hours of culture urokinase-type plasminogen activator (uPA) was detected in COCs and denuded oocytes. In the frozen-thawed COCs and cumulus cell free-oocytes cultured for 24 hours, no PAs were observed. After COCs were cultured for 48 hours, tissue-type plasminogen activator (tPA) and tPA-PAI were observed in COCs only. In the frozen-thawed COCs and cumulus cell free-oocytes cultured for 48 hours, no PAs were observed. These results suggest that uPA, tPA and tPA-PAI are produced by porcine COCs, but only uPA by oocytes during maturation for 24 hours. Only tPA, and tPA-PAI are produced by COCs cultured for 48 hours, and no PAs are produced by denuded-oocytes cultured for 48 hours. In all of the frozen-thawed groups, no PAs are observed by COCs and denuded-oocytes.