본 연구는 소형 개의 불임 해결과 체외수정란을 생산하기 위한 방안의 하나로써 난자의 형태, 번식주기, 배양시간 및 활성화 처리가 난포란의 체외성숙 및 체외발생에 미치는 영향을 조사하였다. 1. 신선, salt 및 4에 보존한 난소로부터 채취한 난구세포부착 난자와 나화 난자로 각각 체외수정시켰을 때 16세포기로의 발생율은 14.3%, 5.0% 및 7.5%, 2.8%, 5.7% 및 0.0%로써 난구세포 부착난자군의 체외발생율이 나화 난자군에 비해 높게 나타
On June 14, 2008 (the first experiment) and July 24, 2008 (the second experiment), the shores of the Boseong River and the sandy beaches, Seokgok-myun, Moksadong-myun, Gokseong-gun in Jeollanam Province were investigated and a total of 29 soft-shelled turtle (Tryonyx sinensis) eggs in the natural spawning nest eggs were collected (13 eggs were collected in the first experiment and 16 eggs in the second experiment). The temperatures in the natural spawning nests were 25.9- 36.9±0.5℃, the depth of the eggs was 5.2-7.5±0.5 cm as the distance of the average 6.4±0.5 cm. 29 eggs were scattered at least 0.2 cm interval. Artificial incubation of 29 eggs was conducted in artificial nest boxes in thermo-plastic composition of the incubator, and then incubated at 26.5-35.5±0.5℃, and an average constant temperature was 31.2-32.1±1.0℃. The incubation days ranged from 53 to 55. In case of most turtles, incubation at 31℃ (higher temperatures) generally produces all or mostly females, while incubation at 25℃(cooler temperatures) produces all or mostly males. Exceptionally, in case of genus Trionyx, the sex ratio of female : male of T. sinensis of a freshwater soft-shelled turtle was approximately 1:1, which differs from other genera of turtles and makes T. sinensis Strauch only turtles presently known to lack temperature-dependent sex determination.
Omija-cheong, concentrated extracts from sugar-treated Omija fruit (Schisandra chinensis Baillon), is produced by traditional manner in Korea. The quality characteristics of Omija-cheong processed at low temperature with a pilot-scale were investigated to optimize the incubation time. With increasing incubation time in processing Omija-cheong, the pH level of Omija-cheong remained constant, while titratable acidity and organic acids increased. Fresh Omija fruits contained citric, malic and succinic acids, most of which were extracted into concentrated extracts after 37 days of incubation and reached to the stable concentration after 47 days of incubation. Titratable acidity in Omija-cheong gradually increased from 1.18% to 2.71%, and also was correlated with total concentration of organic acids. About 80% of supplemented sucrose for manufacturing Omija-cheong was converted into glucose and fructose until 68 days of incubation, and the composition of free sugars was maintained to be stable up to 138 days of incubation. The contents of total flavonoids and phenolic compounds in Omija-cheong were 24.1 mg-GAE/L and 1,635 mg-QE/L at 57 days of incubation, which were more than 9 and 5 times higher than those in Omija fruits, respectively. From the quality characteristics in processing Omija-cheong by low-temperature incubation, more than 60 days of incubation is required for the constant quality and value-added beverage.
Heat shock pretreatment, dark culture period and washing medium could have marked effects on microspore embryogenesis. A heat shock pretreatment of microspores at 32.5°C for 48 hours gave high production rate of microspore-derived embryo (MDE) when compare to shorter and longer period. The yield of MDE increased significantly when microspore cultured for 15 days at 25℃ in dark condition followed by heat shock pretreatment. MDE were browned and lost vitality when dark treatment period extended longer than 15 days. This is caused by an insufficient oxygen and light for growing embryo which already formed during dark treatment period. The vitality of a microspore isolated from flower bud stored at 4℃ become decreased at the very first day and the vitality of microspore stored at 4℃ in the form of flower bud itself become decreased from the 5th day after storage. This shows the possibility of getting a certain period of storage for a suitable flower bud in MDE formation. The yield of MDE was most effective when isolated microspore was had with MS medium compared to B-5 and NLN medium and also showed most effective result with sucrose 130 gL-1 in additional sucrose concentration. The above result is thought to be very useful for the development of a new cultivar of radish and other many crops including Brassica using haploid breeding technology.
This study was conducted to develop an efficient cryopreservation method of human embryonic stem (ES) cells using vitrification. In an initial experiment, sub-clumps of human ES cells (CHA-hES3 and CHA-hES4) were vitrified using grids after incubation with STO feeder cells for 1 or 16 h (Groups 1-1 and 1-2, respectively). After storage for months, thawed clumps were re-plated on a fresh feeder layer. The survival rates of warmed CHA-hES3 and CHA-hES4 cells of Group 1-2 were significantly higher than those of the corresponding Group 1-1 cells. In the second experiment, human ES cells were vitrified after incubation with feeder or feeder-conditioned medium (Groups 2-1 to -7). Relative mRNA expression of BM proteins and survival rates were increased following incubation of ES cells with fresh feeder cells for 16 h. In conclusion, increasing of tight adhesion between ES cells by extended incubation with feeder could reduce cryoinjury after vitrifying/warming.