Several cloned animals have been produced using somatic cell nuclear transfer (SCNT) and have interested in producing the transgenic cloned animals to date. But still its efficiency was low due to a number of reasons, such as sub-optimal culture condition, aberrant gene expression and nuclear reprogramming. The purpose of this study was to analyze gene expression pattern in in vitro fertilized (IVF) or SCNT pre-implantation embryos. IVF- or SCNT-embryos were cultured in media supplemented with different proteins (FBS and BSA) or energy sources (glucose or fructose). Blastocysts from IVF or SCNT were analyzed using semi-quantitative RT-PCR in terms of developmentor metabolic-related genes. Culture medium supplemented different proteins or energy sources had affected on the expression of developmental or metabolic genes in the SCNT blastocysts.

Substantial efforts have been made to manipulate ruminal environment in a hope to enhance ruminal fermentation efficiency for better ruminant productivity. Some of examples are methane inhibitors, antibiotics, microbial enzymes, fatty acids and/or lipid feeding, buffering agents, ionophores and probiotics. Of these efforts, the non-ionic surfactant (NIS) has been known for its stimulation to release enzymes from a range of anaerobic microbes. A series of studies were conducted 1) to evaluate NIS diluted with water and ethanol on in vitro ruminal fermentation and 2) to determine the influence of diluted NIS on digestibility of feedstuffs. In first experiment (Exp. 1), NIS was diluted with water or ethanol to measure its effects on in vitro microbial growth, ruminal enzyme activities and gas production by mixed ruminal microbial culture. The NIS was diluted with water or ethanol separately in a 1:5 ratio (w/v). Water and ethanol-diluted NIS with wheat flour were added with rice straw-based mixed ruminal microbial cultures at the rate of 2 ㎎ NIS/16 ㎖ McDougall buffer plus 4 ml ruminal fluid solution. The mixed ruminal microbial culture was run without any NIS addition as control. Addition of NIS either diluted with water or ethanol has significantly reduced the gas production in mixed ruminal microbial culture at 12 and 24 h of incubation. At 48 h post incubation, gas production was the highest with the addition of ethanol diluted NIS followed by water-diluted NIS and control. Carboxy methyl cellulase activity in rice straw-based mixed ruminal bacterial culture was significantly higher with the addition of ethanol-diluted NIS compared with the addition of water-diluted NIS and control at 24 and 72 h post incubation. In second in vitro experiment (Exp. 2), effects of addition of ethanol diluted NIS on dry matter (DM) digestibility of alfalfa hay, gas production, pH and cellular growth in mixed ruminal microbial culture were examined. Alfalfa hay based mixed ruminal microbial culture without any NIS addition was run as a control. The pH of mixed ruminal microbial culture was significantly lower than control at all post incubation sampling hours. In vitro DM digestibility of alfalfa hay was significantly higher with the addition of NIS compared with control. Gas production was significantly less with NIS addition compared with control at all post incubation sampling hours. Microbial growth in mixed ruminal microbial culture was significantly increased with the addition of NIS compared to control.

본 연구는 기내에서 재생된 오리엔탈나리 ‘Casa Blanca’ 와 ‘Siberia’ 소인경의 새로운 휴면 타파방법을 모색하고 자 생장조절물질을 첨가한 온탕처리 조건을 검토하였다. 오리엔탈나리 ‘Casa Blanca’와 ‘Siberia’ 두 품종 모두 30oC 증류수, 120분 온탕처리에서 맹아소요일수는 단축되 었으나 맹아율이 낮았다. GA4+7 100mg·L−1용액을 35oC 에서 1시간 온탕 처리에 의해서 ‘Casa Blanca’와 ‘Siberia’의 맹아율은 각각 76.7%으로 크게 촉진되었고 정식 2년차의 생장과 구근 비대가 촉진되었으며, 휴면에 따른 호흡정도를 반영하는 미토콘드리아 단백질 함량과 fumarase 활성도 높아 휴면타파의 생리적 원인도 구명 하였다. 생장조절물질(GA4+7)을 첨가한 온탕처리에 의해 기내에서 재생된 오리엔탈나리 소인경의 휴면을 효과 적으로 타파할 수 있었다.

본 실험은 Dendrobium nobile 계통의 교잡종인 'Hamana Lake Dream' X 'No. 55' 유묘의 기내삽목 시 증 식 및 생육에 알맞은 배지와 배양조건을 구명하고자 실시하였다. Hyponex배지보다 MS배지에서 생장이 잘되었는데 MS배지에 활성탄을 0.1과 1.0g/L 수준에서 첨가했을 때 줄기수와 엽장, 근수, 생체중이 양호 하였으나 2.0g/L이상에서는 감소하였으나 통계적 유의성은 없었다. MS배지에 agar, sucrose, peptone, gelite 를 농도별로 첨가한 실험에서는 agar 5g/L 첨가 시 줄기 길이와 엽폭, 생체중의 값이 높았고, 절간장과 근수는 sucrose 40g/L을 첨가했을 때 효과적이었다. 또한 줄기수와 엽수는 peptone 1g/L에서 효과적이었으 나 gelite는 모든 농도에서 대조구에 비해 저조하였다. 기내삽목 시 배양온도는 28℃가 알맞았고 동일한 온도조건에서는 MS배지가 Hyponex배지보다 더 효과적이었다. 또한 sucrose 농도별 실험에서는 28℃에서 40 g/L를 첨가하는 것이 좋았다.

A fiber fraction (Aloe cellulose), the by-product obtained from Aloe vera gel processing was freeze dried and investigated for in vitro glucose/ bile acid retarding effects of powdered sample (100 mesh) comparing with commercial α-cellulose as a reference sample. We also examined the effectiveness of physiological functionality such as the antiobesity and anti-constipation on Sprague-Dawley (SD) rat. The Aloe cellulose powders during in vitro dialysis experiment for 2 hours exhibited the glucose and bile acid retarding index of 20.32-35.2% and 53.13-28.30%, respectively. Especially, freeze dried aloe cellulose showed the 2.5 and 1.2-6 times higher effect on in vitro glucose and bile acid retardation than those of α-cellulose. These relatively good retarding effects on glucose and bile acid diffusion suggest a potential of preventing from diabetes and arteriosclerosis of some extent. Also, the results from animal experiments on SD rats fed a high-fat diet for 4 weeks suggested that Aloe cellulose might be used as a novel dietary fiber showing an effective anti-obesity and anti-constipation effect.

The purpose of this study was to investigate the effects of the collection time, co-culture and sperm penetration of canine oocytes on in vitro maturation and fertilization. The oocytes were cultured in TCM-199 media containing hormonal supplements (10% FCS, 10 IU/ml HCG, 10 IU/ml PMSG) at 5% CO2, 95% air, 38℃. The in vitro maturation rate to MⅡ stage of in vitro oocytes recovered from ovaries that collected at follicular, luteal and inactive phases of the reproductive phase for 44～72 hrs were 19.2%, 12.2%, and 6.0%, respectively. Follicular phases oocytes had a significantly higher in vitro maturation rate than oocytes collected at luteal and anestrus stage (p<0.05). The in vitro maturation rates to the MII stage of canine oocytes after 48 hrs of culture with glutathione, pyruvate, or glutathione + pyruvate were 12.5%, 10.7%, and 17.5%, respectively. This was higher than that in both alone or the combination of the two compared to the control group (19.0%). The sperm penetration rates of in vitro matured oocytes by fresh and frozen semen were 29/80 (36.3%) and 18/80 (22.5%), respectively. Although there are limited reports about canine oocytes co-culture and in vitro fertilization, our results on in vitro maturation is comparable to the results from other researches.

Human fibroblasts that maintain the structural integrity of connective tissues by secreting precursors of the extracellular matrix are typically cultured with serum. However, there are potential disadvantages of the use of serum including unnatural interactions between the cells and the potential for exposure to animal pathogens. To prevent the possible influence of serum on fibroblast cultures, we devised a serum-free growth method and present in vitro data that demonstrate its suitability for growing porcine fetal fibroblasts. These cells were grown under four different culture conditions: no serum (negative control), 10% fetal bovine serum (FBS, positive control), 10% knockout serum replacement (KSR) and 20% KSR in the medium. The proliferation rates and viabilities of the cells were investigated by counting the number of cells and trypan blue staining, respectively. The 10% FBS group showed the largest increase in the total number of cells (1.09 × 105 eell₃/ml). In terms of the rate of viable cells, the results from the KSR supplementation groups (20% KSR:64.7%; 10% KSR: 80.6%) were similar to those from the 10% FBS group (68.5%). Moreover, supplementation with either 10% (30 × 104 eell₃/ml) or 20% KSR (4.8 × 104 cells/ml) produced similar cell growth rates. In conclusion, although KSR supplementation produces a lower cell proliferation rate than FBS, this growth condition is more effective for obtaining an appropriate number of viable porcine fetal fibroblasts in culture. Using KSR in fibroblast culture medium is thus a viable alternative to FBS.