• Journal of Biosystems Engineering
• /
• v.41 no.1
• /
• pp.60-65
• /
• 2016

Purpose: The necessity for precise manipulation of bioparticles has greatly increased in the fields of bioscience, biomedical, and environmental monitoring. Dielectrophoresis (DEP) is considered to be an ideal technique to manipulate bioparticles. The objective of this study is to develop a DEP microfluidic device that can trap fluorescent beads, which mimic bioparticles, at the low voltage and frequency of the sinusoidal signal supplied to the microfluidic device. Methods: A DEP microfluidic device, which is composed of polydimethylsiloxane (PDMS) channels and interdigitated electrode networks, is fabricated to trap fluorescent beads. The geometry of the interdigitated electrodes is determined through computational simulation. To determine the optimum voltage and frequency of the sinusoidal signal supplied to the device, the experiments of trapping beads are conducted at various combinations of voltage and frequency. The performance of the DEP microfluidic device is evaluated by investigating the correlation between fluorescent intensities and bead concentrations. Results: The optimum ratio of the widths between the negative and positive electrodes was 1:4 ($20:80{\mu}m$) at a gap of $20{\mu}m$ between the two electrodes. The DEP electrode networks were fabricated based on this geometry and used for the bead trapping experiments. The optimum voltage and frequency of the supplied signal for trapping fluorescent beads were 15 V and 5 kHz, respectively. The fluorescent intensity of the trapped beads increased linearly as the bead concentration increased. The coefficient of determination ($R^2$) between the fluorescent intensity and the bead concentration was 0.989. Conclusions: It is concluded that the microfluidic device developed in this study is promising for trapping bioparticles, such as a cell or virus, if they are conjugated to beads, and their concentration is quantified.

• 제어로봇시스템학회:학술대회논문집
• /
• 2003.10a
• /
• pp.2451-2456
• /
• 2003

In biological cell manipulation, manual thrust or penetration of an injection pipette into an egg is currently performed by a skilled operator, relying only on visual feedback information. Massive load of various micro injection of either genes, fluid or cells in the postgenomic era calls a more reliable and automatic micro injection system that can test hundreds of genes or cell types at a single experiment. We initiated to study cellular force sensing in zebrafish eggs as the first step for the development of a more controllable micro injection system by any inexperienced operator. Zebrafish eggs at different developmental stages were collected and an integrated biomanipulation system was employed to measure cellular force during penetrating the egg envelope, the chorion. First of all, the biomanipulation system integrated with cellular force sensing instrument is implemented to measure the penetration force of cell membranes and characterize mechanical properties of zebrafish embryo cells. Furthermore, implementation of cellular force sensing system and calibration are presented. Finally, the cellular force sensing of penetrating cell membranes at each developmental stages was experimentally performed. The results demonstrated that the biomanipulation system with force sensing capability can measure cellular force at real-time while the injection operation is undergoing. The magnitude of the measured force was in the range of several hundreds of uN. The precise real-time measurement should provide the first step forwards for the development of an automatic and reliable injection system of various materials into biological cells.

• Proceedings of the Korean Society of Plant Biotechnology Conference
• /
• 2002.04a
• /
• pp.49-58
• /
• 2002

Oxidative stress derived from reactive oxygen species (ROS) is one of the major damaging factors in plants exposed to environmental stress. In order to develop the platform technology to solve the global food and environmental problems in the 21s1 century, we focus on the understanding of the antioxidative mechanism in plant cells, the development of oxidative stress-inducible antioxidant genes, and the development of transgenic plants with enhanced tolerance to stress. In this report, we describe our recent results on industrial transgenic plants by the gene manipulation of antioxidant enzymes. Transgenic tobacco plants expressing both superoxide dismutase (SOD) and ascorbate peroxidase (APX) in chloroplasts were developed and were evaluated their protection effects against stresses, suggesting that simultaneous overexpression of both SOD and APX in chloroplasts has synergistic effects to overcome the oxidative stress under unfavorable environments. Transgenic tobacco plants expressing a human dehydroascorbate reductase gene in chloroplasts were showed the protection against the oxidative stress in plants. Transgenic cucumber plants expressing high level of SOD in fruits were successfully generated to use the functional cosmetic purpose as a plant bioreactor. In addition, we developed a strong oxidative stress-inducible peroxidase promoter, SWPA2 from sweetpotato (Ipomoea batatas). We anticipate that SWPA2 promoter will be biotechnologically useful for the development of transgenic plants with enhanced tolerance to environmental stress and particularly transgenic cell lines engineered to produce key pharmaceutical proteins.

• Plant Biotechnology Reports
• /
• v.4 no.2
• /
• pp.125-128
• /
• 2010

Culture conditions were established for high frequency plant regeneration via somatic embryogenesis from cell suspension cultures of Nymphoides coreana. Zygotic embryos formed pale-yellow globular structures and calluses at a frequency of 85.6% when cultured on half-strength Murashige and Skoog (MS) medium supplemented with 0.3 $mg\;l^{-1}$ of 2,4-D. However, the frequency of pale-yellow globular structures and white callus formation decreased slightly with an increasing concentration of 2,4-D up to 10 $mg\;l^{-1}$ with the frequency rate falling to 16.7%. Cell suspension cultures were established from zygotic embryo-derived calluses using half-strength MS medium supplemented with 0.3 $mg\;l^{-1}$ of 2,4-D. Upon plating onto half-strength MS basal medium, over 92.3% of cell aggregates gave rise to numerous somatic embryos and developed into plantlets. Regenerated plantlets were successfully transplanted into potting soil and achieved full growth to an adult plant in a growth chamber. The high frequency plant regeneration system for Nymphoides coreana established in this study will be useful for genetic manipulation and cryopreservation of this species.

• Journal of Animal Science and Technology
• /
• v.58 no.3
• /
• pp.13.1-13.7
• /
• 2016

Background: In the present study, various freezing containers were tested for mouse embryos of respective developmental stages; embryos were vitrified and then their survival rate and developmental rate were monitored. Mouse two cell, 8 cell, and blastula stage embryos underwent vitrification freezing-thawing and then their recovery rate, survival rate, development rate, and hatching rate were investigated. Methods: EM-grid, OPS, and cryo-loop were utilized for vitrification freezing-thawing of mouse embryos. Results: It was found that recovery rate and survival rate were higher in the group of cryo-loop compared to those of EM-grid (p < 0.05). Embryonic development rate, two cell embryos to blastocyst, as well as hatching rate were higher in the control group compared to the EM-grid group and OPS group (p < 0.05), yet no difference was noted between the control group and cryo-loop group. Development rate and hatching rate of eight cell morulae and blastocysts were all lower in the treatment groups than the control group whilst hatching rate of blastocysts was higher in the control group compared to the groups of EM-grid and OPS (p < 0.05); although the cryo-loop group was shown to be slightly higher than other groups, it was not statistically significant. Conclusions: In the study, we investigate effects of freezing containers on vitrified embryos of respective developmental stages; it was demonstrated that higher developmental rate was shown in more progressed (or developed) embryos with more blastomeres. There was however, no difference in embryonic development rate was shown amongst containers. Taken together, further additional studies are warranted with regards to 1) manipulation techniques of embryos for various vitrification freezing containers and 2) preventive measures against contamination via liquid nitrogen.

• Archives of Plastic Surgery
• /
• v.39 no.5
• /
• pp.518-521
• /
• 2012

Background Skin cancer is the most common malignant tumor in humans. Basal cell carcinoma (BCC) and squamous cell carcinoma (SCC) are the two most common types of skin cancers. When skin cancer is clinically suspected, preoperative biopsies are recommended for a definite diagnosis. However, despite a concern over potential increased risk of metastasis associated with mechanical manipulation, there have been few investigations into whether a preoperative biopsy affected the recurrence of BCC and SCC. Methods Primary BCC or SCC patients who underwent standard surgical excision from 1991 to 2010 were reviewed and a retrospective analysis was performed. Ultimately, 45 BCC patients and 54 SCC patients, who did not meet the exclusion criteria, were analyzed. To identify whether a preoperative biopsy affected the recurrence of BCC and SCC, the recurrence rates of each with and without biopsy were compared. Results Preoperative biopsy had no statistically significant effect on recurrence (BCC, P=0.8680; SCC, P=0.7520). Also, there was no statistical significance between the interval from initial biopsy to first operation and recurrence (BCC, P=0.2329; SCC, P=0.7140). Even though there was no statistical significance, the mean interval from the biopsy to the operation among the BCC patients who underwent preoperative biopsy was 9.2 months in those who had recurrence and 2.0 months in those who had no recurrence. Conclusions There was no statistically significant relationship between preoperative biopsy and recurrence of BCC and SCC. However, there was a tendency toward recurrence in patients with a longer interval between the biopsy and the corrective operation in BCC.

• Molecules and Cells
• /
• v.39 no.4
• /
• pp.330-336
• /
• 2016

Long non-coding RNAs (lncRNAs) are involved in multiple cellular events, as well as in tumorigenesis. Colon cance-rassociated transcript-1 (CCAT1) gene encodes an lncRNA whose over-activation was observed in an expanding list of primary human solid tumors and tumor cell lines, however its biological roles in acute myeloid leukaemia (AML) has not been reported yet at present. In this study, the aberrant upregulation of CCAT1 was detected in French-American-British M4 and M5 subtypes of adult AML patients. By gain- and loss-of-function analysis, we determined that CCAT1 repressed monocytic differentiation and promoted cell growth of HL-60 by sequestering tumor suppressive miR-155. Accordingly, a significant decrease in miR-155 level was detected in AML patients. Reintroduction of miR-155 into HL-60 cells restored monocytic maturation and repressed cell proliferation. Furthermore, CCAT1 could up-regulated c-Myc via its competing endogenous RNA (ceRNA) activity on miR-155. In conclusion, these results revealed new mechanism of lncRNA CCAT1 in AML development, and suggested that the manipulation of CCAT1 expression could serve as a potential strategy in AML therapy.

• Journal of Embryo Transfer
• /
• v.9 no.3
• /
• pp.229-234
• /
• 1994

This experiment was carried out to develop the model system for mass production of biomedical and nutritional proteins (human proteins) through mamraary gland of the transgenic cattle produced by gene manipulation and embryological technologies. Human growth hormone gene fused with rat $\beta$-casein gene promoter was microinjected into pronuclei of one cell bovine embryos produced by in vitro fertilization. After microinjection, embryos were cultured in vitro for 6 or 7 days. Twenty embryos reaching to blastocysts were transferred to 10 beef recipients, each receiving two embryos. Recipients were diagnosed for pregnancy by rectal palpation at 76 days after embryo transfer. One of them was pregnant to term and produced a female calf weighing 21 kg at 280 days following embryo transfer. DNA was extracted from umbilical cord tissue and blood of calf born for confirming gene insertion. As determined by Southern hybridization, the transgene was not found.

• Transactions of the Korean Society of Mechanical Engineers B
• /
• v.32 no.1
• /
• pp.62-69
• /
• 2008

A post-processing program based on the OOP(Object-Oriented Programming) concept has been developed for flow visualization of the flow analysis code(PowerCFD) using unstructured cell-centered method. User-friendly GUI(GTaphic User Interface) has been built on the base of MFC(Microsoft Foundation Class). The program is organized as modules by classes including those based on VTK(Visualization ToolKit)-library, and these classes are made to function through inheritance and cooperation which is an important and valuable OOP concept. The major functions of this post-processor program are introduced and demonstrated, which include mesh plot, contour plot, vector plot, surface plots, cut plot, clip plot, xy-plot and streamline plot as well as view manipulation (translation, rotation, scaling etc).

• Asian-Australasian Journal of Animal Sciences
• /
• v.7 no.4
• /
• pp.459-466
• /
• 1994

Primordial germ cells (PGCs) in aves are the progenitor cells for the gametes. These cells first appear in the epiblast (Eyal-Giladi et al.. 1981). Then translocate and concentrate to endoderm of germinal crescent area in the junction of the area opaca and area pellucida lateral to the primitive streak in stage 4 through 7. They separate from the endoderm, temporarily circulate via the blood vascular system, leave the blood vessels, and finally settle down in the gonadal anlagen at stage 20-24 where they rapidly proliferate to form germ cells. Recently, several attempts have been made to introduce foreign gene into the avian genome to form a transgenic chicken. The stem cells most readily available as vehicles for genetic manipulation of germline in avian species are the PGCs. PGCs have recently been manipulated genetically and used successfully as a vector for gene transfer.