Spermatogonial stem cells(SSCs) only are responsible for the generation of progeny and for the transmission of genetic information to the next generation in male. Other in vitro studies have cultured SSCs for proliferation, differentiation, and genetic modification in mouse and rat. Currently, information regarding in vitro culture of porcine Germline Stem Cell(GSC) such as gonocyte or SSC is limited and is in need of further studies. Therefore, in this study, we report development of a successful culture system for gonocytes of neonatal porcine testes. Testis cells were extracted from 10～14-day-old pigs. These cells were harvested using enzymatic digestion, and the harvested cells were purified with combination of percoll, laminin, and gelatin selection techniques. The most effective culture system of porcine gonocytes was established through trial experiments which made a comparison between different feeder cells, medium, serum concentrations, temperatures, and O2 tensions. Taken together, the optimal condition was established using C166 or Mouse Embryonic Fibroblast(MEF) feeder cell, Rat Serum Free Medium(RSFM), 0% serum concentration, 37℃ temperature, and O2 20% tension. Although we discovered the optimal culture condition for proliferation of porcine gonocytes, the gonocyte colonies ceased to expand after one month. These results suggest inadequate acquirement of ingredients essential for long term culture of porcine GSCs. Consequently, further study should be conducted to establish a successful long-term culture system for porcine GSCs by introducing various growth factors or nutrients.

The current study was designed to evaluate the effects of the reactive oxygen species (ROS) generated with a xanthine (X) and xanthine oxidase system (XO) on sperm function and DNA fragmentation in porcine spermatozoa. ROS were produced by a combination of 1,000 μM X and 50 mU/ml XO. The ROS scavengers such as superoxide dismutase (SOD) (200 U/ml) and catalase (CAT) (500 U/ml) were also tested. Spermatozoa were incubated for 2 hours in BWW medium with a combination of X-XO supplemented with or without antioxidants at 37℃ under 5% CO2 incubator. Ca-ionophore-induced acrosome reaction, the proportion of swollen spermatozoa under hypo-osmotic condition, malondialdehyde formation for the analysis of lipid peroxidation, and the proportion of DNA fragmentation were determined after 2 hours incubation. The action of ROS on porcine spermatozoa resulted in decreased Ca-ionophore-induced acrosome reaction and membrane integrity, increased the formation of malondialdehyde, and the proportion of sperm with DNA fragmentation(p<0.05). The toxic effects caused by ROS were completely alleviated by CAT in terms of sperm function and characteristics, however SOD did not serve the same scavenger effect as CAT. To conclude, the ROS can cause significant damage to porcine sperm functions and characteristics, which can be minimized by the use of antioxidants.

Petroleum pitch and coke with wet mixture method or with dry mixture method were investigated to develop the composite anodic carbon material of high power lithium ion battery. Cokes coated with pitch were obtained by the heat treatment of mixture of cokes and pitch with different weight ratios at 800~1200℃. The charge and discharge characteristic of the consequent composite anodic carbon material assembled in batteries was tested. Cokes with wet mixture method have a smooth surface and their capacity changed little with changing temperature and content as compared to the cokes with dry mixture method. Although the reversible capacities showed different values by the anode manufacturing method, the composite anode with the mixture of 20 wt% of petroleum pitch and 80 wt% of coke showed the higher power capability and initial efficiency than the pitch based anode. However, the reversible capacity of the composite anode showed the reduced value as compared with the pitch based anode.

본 연구는 돼지에 있어서 정원줄기세포를 포함하는 정소세포를 recipient 돼지의 정소 내로 이식할 수 있는 기법을 개발하기 위하여 시행되었다. 공여세포는 10~14일령의 돼지로부터 채취된 정소에서 효소처리법을 이용하여 회수하였고, recipient의 정소 내로 이식하기 전 형광 마커(PHK26)로 표지하였다. 외과적 수술을 통하여 recipient 돼지부터 정소를 꺼낸 후 초음파 기기와 이식 장치를 이용하여 형광표지된 공여세포를 recipient 정소의 세정관 내로 이식하였다. 14주령의 recipient 정소에 5~7ml 의 공여 세포부유액을 주입하여 정소 내 50% 이상의 세정관 내로 새포부유액의 주입이 가능하였고, 세포부유액이 주입된 세정관 내에서 형광표지된 정소세포들이 고루 이식되어짐이 관찰되었다. 본 연구에서 개발한 이식 기법을 이용하여 효율적인 정소세포의 이식이 가능함에 따라 향후 돼지 정원줄기세포의 연구 및 활용법 개발에 획기적인 돌파구가 마련될 것으로 기대된다.

Numerical analysis using commercial CFD code was carried out to develop the drag force type vertical axis hydraulic turbine for the improvement of the production efficiency of small hydro energy at low flow velocity condition. Blade pressure changes and internal flows were analyzed according to the presence or absence of the hydraulic turbine blade holes at flow velocity of less than 1.0~3.0 m/s. According to the numerical results, the pressure and flow velocity is severly affected by the flow velocity in turbine blade with no holes, while the influence of flow velocity is comparatively decreased in turbine blade with holes. It is also found that the pressure and flow velocity on the blade surface with holes are evenly distributed with no singular location and it is believed that forming a hole in the blade may be helpful in terms of structural safety.

이 연구의 목적은 개착식 전력구 콘크리트에 발생하는 부등건조수축에 의한 균열특성을 파악하고, 수축저감제 혼입에 따른 건조수 축균열특성을 분석하는데 있다. 따라서 이 연구에서는 전력구 박스형 콘크리트 구조물의 부등건조수축에 의한 균열특성을 파악하기 위하여 세 가지 주요영향인자를 고려한 수치해석과 수치해석을 한 결과와 실제 전력구 타설을 하여 측정한 차이를 확인해 보았다. 본 연구로부터 건조 수축 시험체에 대한 건조수축량을 산정에 대한 해석 기법을 개발하였으며, 실제 전력구 타설 실험을 통해 종방향 및 횡방향 균열발생 가능성을 입증 하였다. 전력구 실증시험체 건조수축량 수치해석 결과, 균열저감 콘크리트 배합의 경우 종방향 및 횡방향 응력에 있어 일반배합에 비해 40~50%정도의 현저한 감소를 보이며, 이에 따라 균열저감 콘크리트를 전력구에 적용하였을 경우 현저한 균열감소 효과를 보일 것으로 예측된다.

Recently, interests for maintenance of transmission tower are increasing to extend life of structures and reduce maintenance cost. However, existing classical diagnosis method of corrosion deteriorated degree on the transmission tower steel members, visual inspection, has a problem that error often due to difference of inspector’s individual knowledge and experience. In order to solve the problem, this study carried out to develop the corrosion deterioration inspection tool for transmission tower steel members. This tool is composed of camera equipment and computer-aided diagnosis system. We standardized the photographing method by camera equipment to obtain suitable pictures for image processing. Diagnosis system was designed to evaluate automatically degree of corrosion deterioration for member of transmission tower on the basis of the RGB color image processing techniques. It is anticipated that developed the corrosion deterioration inspection tool will be very helpful in decision of optimal maintenance time for transmission tower corrosion.

이 연구의 목적은 개착식 전력구 콘크리트에 발생하는 부등건조수축에 의한 균열특성을 파악하고, 그 제어방법을 제시하는데 있 다. 건조수축균열은 콘크리트 내부의 수분확산계수의 영향을 크게 받으며, 수분확산계수는 콘크리트 내부에서의 수분이동속도를 결정하는 주요인자이다. 수분확산계수와 더불어 콘크리트 표면의 표면계수와 외부의 상대습도는 콘크리트 내부에서 외부로의 수분이동에 영향을 미친 다. 따라서 이 연구에서는 전력구 박스형 콘크리트 구조물의 부등건조수축에 의한 균열특성을 파악하기 위하여 세 가지 주요영향인자를 고려 한 수치해석을 수행하였다. 수치해석 결과, 수분확산계수와 표면계수가 증가할수록 상부슬래브의 부등건조수축에 의한 균열발생시점이 빨라 지며, 세 가지 요인 중에 콘크리트의 부등건조수축에 의한 균열발생 특성에 가장 큰 영향을 미치는 것은 외기습도인 것으로 나타났다. 이 연구 결과를 분석한 결과, 개착식 전력구 시공시에 콘크리트 타설 후 표면보습이나 살수양생과 같이 외기습도를 증가시키는 것이 부등건조수축에 의한 균열제어에 가장 효과적인 것으로 판단되며, 콘크리트 재료적 측면의 균열저감방법으로 수분확산계수와 표면계수를 결정하는 콘크리트 의 배합이나 재료특성을 적절히 선정함으로써 균열의 진전속도나 발생시점을 제어할 수 있을 것으로 판단된다.

The aim of this study was to enhance the proliferation efficiency of spermatogonial stem cells (SSCs). In order to improve the proliferation efficiency, we investigated new factors that promote the proliferation of SSCs using in vitro culture method with natural plant extracts. Germ cell populations containing SSCs were collected 6- to 8-days-old from C57BL/6-TG-EGFP (C57GFP) mice and SSCs were isolated from the collected cells via magnetic-activated cell sorting (MACS). Since then, SSCs were cultured for a week with culture medium containing natural plant extracts at concentration of 0.1, 1, and 10 μg/mL. After a week of culture, we looked for an increase, especially a dose-dependent increase, in the number of cells compared to that of the control group. A dose-dependent increase, in the number of cells was observed in the Petasides japonicus-treated groups. Furthermore, we carried out repeated experiment that is process consisting of selection and additional segmentation to explore new factors for activating SSCs at the molecular level. As a results, Petasides japonicus butanol fraction significantly increased the proliferation rate of SSCs in a dose-dependent manner among Petasides japonicus fraction samples. We identified normal expression level of PLZF in SSCs cultured with plant extracts using immunocytochemistry method. Furthermore, we also carried out qRT-PCR and identified normal expression level of Lhx1 and GFRα1. The finding of this study could contribute to improvement of proliferation and activation for SSCs, using culture method with natural plant extracts.

Spermatogonial stem cells (SSCs) possess the unique capacity of self-renewal and differentiation and thereby can transmit genetic information to the next generation. Combination with techniques such as isolation, culture, preservation, and transplantation of the SSC has facilitated the efficient method for production of transgenic animals, and preservation of livestock and endangered species. The purpose of this study was to genetically modify enriched populations of pre-pubertal germ cells using lentiviral transduction and to develop an efficient in vitro culture system and cryopreservation technique for bovine SSCs. To maximize the efficiency of genetic modification of bovine SSCs, effective enrichment techniques need to be developed. Selection of bovine SSCs using a combination of laminin and gelatin was resulted in a 8-fold enrichment. Selected cells were then transduced using a lentiviral vector containing the transgene for the enhanced green fluorescent protein. Transduction efficiency was 17%. Next, to enhance the efficiency of proliferation for in vitro culture, the effects of various culture conditions and growth factors on bovine cell proliferation were evaluated. Based on the results, we developed the optimal culture conditions [2× rat sertum free medium (rSFM) containing 0.1% FBS together with GDNF, GFRα1, bFGF, EGF, LIF, and CSF-1] for maintaining bovine SSCs over 3 months without any alteration of stem cell characteristics and functions. Also, to develop an effective cryopreservation technique for bovine SSCs, the effects of different freezing methods and various cryoprotective agents were tested. The recovery rate, and proliferation capacity of bovine SSCs were significantly greater for germ cells frozen using tissue freezing methods compared to cell freezing methods. Cryopreservation in the presence of 200 mM trehalose resulted in significantly greater recovery rate, and proliferation capacity of germ cells compared to control. As a results, cryopreservation using tissue freezing methods in the presence of 200 mM trehalose is an efficient cryopreservation protocol for bovine SSCs. Collectively, these findings can serve as a model for comprehensively understanding the biology of SSCs and the factors that regulate male fertility. Furthermore, results of this study will be integral for the continued refinement of techniques to manipulate bovine SSCs.