• Reproductive and Developmental Biology
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• 제33권2호
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• pp.113-117
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• 2009

Animals produced by somatic cell nuclear transfer (SCNT) using genetically modified cells are almost always transgenic, implying that this method is more efficient than the traditional pronuclear microinjection method. Most somatic cells for SCNT in animals are fetus-derived primary cells and successful gene integration in somatic cells will depend on transfection condition. The objective of this study is to evaluate the efficiency of electroporation (Microporator) and liposome reagents (F-6, F-HD, W-EX, W-Q, W-M) for tissue-type plasminogen activator (tPA) gene transfection and to estimate the overall efficiency of transfection of Korean native pig fetal fibroblast cells (KNPFF). Electroporation showed significantly higher transfection efficiency than liposome reagents with regard to the transfection of in vitro cultures in the early stages of development (41.7% with Microporator vs. 18.3% with F-6, 20.0% with F-HD 18.5% with W-EX, 5.0% with W-M and 6.3% W-Q,). Colonies identified as tPA-positives were treated once more with G418 for 10 to 14 days and growing colonies were selected again. When the cells of newly selected colonies were subjected to single-cell PCR, reselection of colonies following second round of G418 selection increased the rate of transgene integration per each colony. These results suggest that transfection with electroporation is the most efficient and the second rounds of G418 selection may be an effective method for transfection of porcine fetal fibroblast cells.

• Reproductive and Developmental Biology
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• 제30권4호
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• pp.279-285
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• 2006

Vitrification has been suggested to be an effective method for the cryopreservation of human ES cells. However, the efficiency of vitrification with different vehicles remains a matter of ongoing controversy. The objective of this study was to assess the efficiency of cryopreservation in human ES cells by vitrification using different vehicles. A human ES cell line and a variety of vehicles, including micro-droplet (MD), open-pulled straw (OPS) and electron microscopic grid (EM-grid), were employed in an attempt to assess vitrification efficiency. In order to evaluate the survivability and the undifferentiated state of the post-vitrified human ES cells, we conducted alkaline phosphatase staining and characterization via both RT-PCR and immunofluorescence assays. The survival rates of the post-vitrified human ES cells using MD, OPS and EM-grid were determined to be 61.5%, 66.6% and 53.8%, respectively. There also exist significant differences between slow-freezing and vitrification (p<0.01). However, no significant differences were detected between the vehicle types. Finally, the pluripotency of human ES cells after thawing was verified by teratoma formation. Cryopreservation using vitrification is more effective than slow-freezing, and the efficiency of vehicles proved effective with regard to the preservation of human ES cells.

• Reproductive and Developmental Biology
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• 제35권4호
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• pp.449-455
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• 2011

Induced pluripotent stem (iPS) cells have been generated from mouse and human somatic cells by etopic expression of transcription factors. iPS cells are indistinguishable from ES cells in terms of morphology and stem cell marker expression. Moreover, mouse iPS cells give rise to chimeric mice that are competent for germline transmission. However, mice derived from iPS cells often develop tumors. Furthermore, the low efficiency of iPS cell generation is a big disadvantage for mechanistic studies. Nonviral plasmid.based vectors are free of many of the drawbacks that constrain viral vectors. The histone deacetylase inhibitor valproic acid (VPA) has been shown to improve the efficiency of mouse and human iPS cell generation, and vitamin C (Vc) accelerates gene expression changes and establishment of the fully reprogrammed state. The MEK inhibitor PD0325901 (Stemgent) has been shown to increase the efficiency of the reprogramming of human primary fibroblasts into iPS cells. In this report, we described the generation of mouse iPS cells devoid of exogenous DNA by the simple transient transfection of a nonviral vector carrying 2A-peptide-linked reprogramming factors. We used VPA, Vc, and the MEK inhibitor PD0325901 to increase the reprogramming efficiency. The reprogrammed somatic cells expressed pluripotency markers and formed EBs.

• Reproductive and Developmental Biology
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• 제28권1호
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• pp.7-12
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• 2004

This study was performed to establish the effective culture condition for the establishment of clonal lines from porcine fetal fibroblast cells. Fibroblasts derived from a pig fetus (Day 50) were cultured and passaged two times before use. A single cell was seeded in 96-well plates, cultured in medium supplemented with different concentrations of FBS, catalase or $\beta$-mercaptoethanol ($\beta$ME), and classified by cell size and morphology. Cells were passaged two times into 4-well dish before freezing. The establishment efficiencies were not different among different concentrations of FBS (0.3 to 5.1%). However, population doubling time (PDT) was significantly decreased by increasing the FBS concentration (P＜0.05). The establishment efficiency of $\beta$ME-added group (10.4%) was significantly higher than those of catalase-added and control groups (3.5%, and 3.5%, respectively, p＜0.05), and PDT was significantly decreased (23.6 vs 28.1, and 25.5 h, respectively, p＜0.05). However, catalase did not show a positive effect on the establishment efficiency. Cell size and morphology did not affect the establishment efficiency and PDT of clonal lines. The result of present study shows that the establishment efficiency of clonal cell lines can be enhanced by the culture in media supplemented with 30% FBS and $\beta$ME.

• 한국콘텐츠학회:학술대회논문집
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• 한국콘텐츠학회 2008년도 춘계 종합학술대회 논문집
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• pp.301-305
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• 2008

유기 발광 다이오드(Organic light-emitting diode, OLED)는 저전력 구동, 자체발광, 넓은 시야각, 우수한 고해상도, 풀 칼라, 높은 재현성, 빠른 응답속도, 간편한 제조 공정 등의 장점을 가지고 있으나, 고성능 디스플레이로서 실용화하기 위해서는 아직도 해결되어야 할 과제가 많다. 소자의 저소비전력, 제조공정의 안정성, 대형 기판기술, 봉지 기술, 소자의 수명, 풀 컬러화를 위한 적색, 청색, 발광소자의 고휘도 등이 시급하다. 무엇보다 중요한 것은 유기 발광 소자의 효율을 향상시키는 것이 상용화를 위한 키(key) 이다. 이를 위해서 유기 발광 소자의 구조 개선과 새로운 유기 물질 적용을 통해 구동전압을 낮춤으로써 효율을 향상시킬 수 있다. 따라서 본 연구에서는 유기 발광 소자의 효율을 향상시킬 목적으로 ITO/TPD/Znq2+DCJTB/Znq2/Al의 구조와 ITO/CuPc/N PB/Alq3+DCJTB/Alq3/Al의 구조를 가지는 소자의 발광층에 형광염료를 도포한 적색 발광 소자를 제작하고, 그 전기적 및 광학적인 특성을 평가하였다.

• 제어로봇시스템학회:학술대회논문집
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• 제어로봇시스템학회 2003년도 ICCAS
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• pp.215-219
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• 2003

In this paper, we generate a trajectory minimized the energy gait of a biped robot for walking a staircase using genetic algorithms and apply to the computed torque controller for the stable dynamic biped locomotion. In the saggital plane, a 6 degree of freedom biped robot that model consists of seven links is used. In order to minimize the total energy efficiency, the Real-Coded Genetic Algorithm (RCGA) is used. Operators of genetic algorithms are composed of a reproduction, crossover and mutation. In order to approximate the walking gait, the each joint angle is defined as a 4-th order polynomial of which coefficients are chromosomes. Constraints are divided into equality and inequality. Firstly, equality constraints consist of position conditions at the end of stride period and each joint angle and angular velocity condition for periodic walking. On the other hand, inequality constraints include the knee joint conditions, the zero moment point conditions for the x-direction and the tip conditions of swing leg during the period of a stride for walking a staircase.

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2004년도 춘계학술발표대회
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• pp.189-189
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• 2004

Histone acetylation as epigenetic marker plays a critical role in gene expression through the interaction of nucleosomes with DNA, modulating the efficiency which RNA-polymerase can interact with promotors to initiate transcription. After fertilization, highly acetylated chromatin takes place and maintain during 1cell stages. The hyperacetylation may lead minor genome activation for survival and cleavage, and then may affect embryonic genome activation and development to balstocyst. (omitted)

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2002년도 춘계학술발표대회 발표논문초록집
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• pp.10-10
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• 2002

The low efficiency of animal production by nuclear transfer technique is considered to be result of an incomplete reprogramming of the donor cell nucleus, which leads to a lack of, or abnormal expression of developmentally important genes. There are a lot of genes related to embryo development and some of these genes are regulated by imprinting. IGF2 (insulin like growth factor 2) and IGF2R (IGF2 receptor) that play important roles in preimplantation development are included in imprinted genes also. (omitted)

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2002년도 춘계학술발표대회 발표논문초록집
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• pp.66-66
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• 2002

The efficiency of animal production using cloning technology is relatively low. It is considered that the nuclear transferred (NT) embryos proceed inappropriate reconstruction with donor-recipient cell, which lead to a abnormal embryo development, and differential expression of mRNA transcript. Especially, the expression of mRNA on peri-implantation stage embryos is very important factor to decide success of implantation and ongoing pregnancy. (omitted)

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2002년도 춘계학술발표대회 발표논문초록집
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• pp.23-23
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• 2002

In vitro matured porcine oocytes have very small volume of perivitellinespace(PVS). In these respect, the effects of sucrose and polybrene on the efficiency of gene transfer were investigated. As a gene (hGH) transfer vehicle, Vesicular stomatitis virus glycoprotein pseudotyped retroviral vector (VSV-G) was used. Sucrose treatment have no detrimental effect on the rates of cleavage and following development and induced the enlargement of PVS resulting the efficient introduction of retroviral vector stocks into PVS. (omitted)