The purpose of this study was to investigate the development of bovine nuclear transfer (NT) embryos cultured in serum-free conditions. Bovine NT embryos cultured in various culture conditions were compared blastocyst development, total cell number and apoptosis using TUNEL assay. In experiment 1, blastocyst rates of NT embryos were significantly higher (P<0.01) in FBS (22.0％) and BSA (26.6％) groups than in PVA (6.3％) group. Total cell number was significantly higher in FBS (78.4±19.4) and BSA (90.9±29.1) groups than in PVA group (46.0±0.0). Apoptotic cell number was significantly fewer in FBS (3.1±1.4) and BSA (1.7±1.4) groups than in PVA group (7.0±20.0) However, all of results were not different between the FBS and BSA group. In experiment 2, blastocyst rates of NT embryos were significantly higher (P<0.05) in fatty acid free-BSA (FAF-BSA) group (26.8％) than in fraction V-BSA group (11.2％). Total cell number were somewhat higher in FAF-BSA group (89.8±30.7) than in fraction V-BSA group (88.1±19.3). Apoptotic cell number were somewhat fewer in FAF-BSA (1.7±1.5) group than in fraction V-BSA group (4.2±2.9). These findings suggest that serum free condition were effective for the in vitro development of bovine NT embryos. Therefore, we concluded that fatty acid free-BSA has beneficial effect in development bovine NT embryos and can be use as a serum substitute.

The electrical, mechanical and optical capabilities have been tested of the microdensitometer PDS 1010GMS at the Korea Astronomy Observatory. The highest stage of scan speed 255 csu (conventional speed unit) is measured to be 47 mm/s. At this speed the position is displaced by 4 μ m to the direction of scanning and the density is underestimated by 0.4 ∼ 0.7 D . Standard deviation in the measured density is proportional to A − 0.46 , where A is the area of scan aperture. The accuracy of position repeatability is ± 1 μ m , and that of density repeatability is ± ( 0.003 ∼ 0.03 ) D . Callier coefficient is determined to be 1.37; the semispecular density is directly proportional to the diffuse density up to 3.5D. Because the logarithmic amplifier has a finite response time, the densities measured at high scan speeds are underestimated to the degree that speeds higher than 200 csu are inadequate for making an accurate astronomical photometry. After power is on, an about 5 hour period of warming is required to stabilize the system electrically and mechanically as well. On the basis of this performance test, we have determined the followings as the optimum scan parameters for the astronomical photometry: For the scan aperture 10 ∼ 20 μ m is optimal, and for the scan speed. 20 ∼ 50 csu is appropriate. These parameter values are chosen in such a way that they may keep the density repeatability within ± 0.01 D , the position displacement under 1 μ m , and the density underestimation below 0.1D even in high density regions.