• Title/Summary/Keyword: Cell object

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In Vitro Development of Interspecies Nuclear Transfer Embryos using Porcine Oocytes with Goat and Rabbit Somatic Cells

  • Quan, Yan Shi;Naruse, Kenji;Choi, Su-Min;Kim, Myung-Youn;Han, Rong-Xun;Park, Chang-Sik;Jin, Dong-Il
    • Reproductive and Developmental Biology
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    • v.32 no.4
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    • pp.249-253
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    • 2008
  • Interspecies somatic cell nuclear transfer (iSCNT) is a valuable tool for studying the interactions between an oocyte and somatic nucleus. The object of this study was to investigate the developmental competence of in vitro-matured porcine oocytes after transfer of the somatic cell nuclei of 2 different species (goat and rabbit). Porcine cumulus oocytes were obtained from the follicles of ovaries and matured in TCM-199. The reconstructed embryos were electrically fused with 2 DC pulses of 1.1kV/cm for $30{\mu}s$ 0.3M mannitol medium. The activated cloned embryos were cultured in porcine zygote medium-3 (PZM-3), mSOF or RDH medium for 7 days. The blastocyst formation rate of the embryos reconstructed from goat or rabbit fetal fibroblasts was significantly lower than that of the embryos reconstructed from porcine fetal fibroblast cells. However, a significantly higher number of embryos reconstructed from goat or rabbit fetal fibroblasts cultured in mSOF or RDH, respectively, developed to the morular stage than those cultured in PZM-3. These results suggest that goat and bovine fetal fibroblasts were less efficacious than porcine-porcine cloned embryos and that culture condition could be an important factor in iSCNT. The lower developmental potential of goat-porcine and porcine-bovine cloned embryos may be due to incompatibility between the porcine oocyte cytoplasm and goat and bovine somatic nuclei.

Expression and Significance of Hypoxia Inducible Factor-1α and Lysyl Oxidase in Non-small Cell Lung Cancer

  • Ping, Wei;Jiang, Wen-Yang;Chen, Wen-Shu;Sun, Wei;Fu, Xiang-Ning
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.6
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    • pp.3613-3618
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    • 2013
  • Object: To detect expression of hypoxia inducible factor-$1{\alpha}$ (HIF-$1{\alpha}$) and lysyl oxidase (LOX) in non-small cell lung cancer (NSCLC) and explore their roles in prognosis. Methods: The mRNA levels of HIF-$1{\alpha}$ and LOX were investigated by real-time reverse-transcriptase polymerase chain reaction in 40 cases of tumour and paired normal tissues. In addition, protein expression of HIF-$1{\alpha}$ and LOX was examined by immunohistochemistry in 82 cases of tumour and 45 paired normal tissues. The relationship between HIF-$1{\alpha}$ or LOX and clinicopathologic characteristics, as well as the correlation between HIF-$1{\alpha}$ and LOX, were also examined. Kaplan-Meier survival curves and the log-rank test were used to analyze progression-free survival. Results: HIF-$1{\alpha}$ or LOX mRNA levels in tumor tissues was significantly higher than those in paired normal tissues (p<0.01). Positive HIF-$1{\alpha}$ or LOX protein expression in tumor tissues was noted in 46/82 (56.1%) and 49/82 (59.8%) of the cases, respectively, being significantly higher than those in paired normal tissues (p<0.05). There was significant correlation between the expression of HIF-$1{\alpha}$ or LOX and tumor size, lymph node metastasis and pathological stage (p<0.05). The expression of HIF-$1{\alpha}$ and LOX had a significant inverse impact on survival of patients with NSCLC. Conclusion: HIF-$1{\alpha}$ and LOX may play a pivotal role in the development of NSCLC, and may act in synergy to promote the progression of NSCLC.

Osteogenic Differentiation of Bone Marrow Stem Cell using Bi-phase Alginate Scaffold Including BMP-2 (BMP-2를 함유한 2상 알지네이트 담체를 이용한 골수줄기세포의 골분화)

  • Lim, Hyun-Ju;Kim, Hak-Tae;Oh, Eun-Jung;Kim, Tae-Jung;Ghim, Han-Do;Choi, Jin-Hyun;Chung, Ho-Yun
    • Archives of Plastic Surgery
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    • v.37 no.3
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    • pp.207-212
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    • 2010
  • Purpose: The object of this study is to develop a novel BMP-2 delivery system for continuous osteogenic differentiation and to induce osteogenesis of stem cells using a bi-phase alginate carrier in vitro. Methods: Alginate nanoparticle loaded BMP-2 was prepared by the reverse emulsification-diffusion technique. Physical properties and release profiles of alginate carriers were measured by Instron and ELISA kit, respectively. Cell viability and alkaline phosphate activity of hBMSCs differentiation was also evaluated by MTS and Metra BAP assays, respectively. Results: Optimal concentration for bi-phase alginate carrier was determined as 2 wt% by evaluating mechanical and biological properties, and differentiation of BMSCs for bone regeneration. The 2% bi-phase alginate carrier had the lowest initial and final release ratio. In addition, the 2% bi-phase alginate carrier had a little higher ALP activity than the homogeneous carrier. An improved controlled release profile was obtained by combining alginate hydrogel with lyophilized particles. Conclusion: Bi-phase alginate carrier has many advantages such as biocompatibility and controlled release capability. It is expected to be effective as a scaffold and carrier in bone tissue engineering.

Advanced tube formation assay using human endothelial colony forming cells for in vitro evaluation of angiogenesis

  • Lee, Hyunsook;Kang, Kyu-Tae
    • The Korean Journal of Physiology and Pharmacology
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    • v.22 no.6
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    • pp.705-712
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    • 2018
  • The tube formation assay is a widely used in vitro experiment model to evaluate angiogenic properties by measuring the formation of tubular structures from vascular endothelial cells (ECs). In vitro experimental results are crucial when considered the advisability of moving forward to in vivo studies. Thus, the additional attentions to the in vitro assay is necessary to improve the quality of the pre-clinical data, leading to better decision-making for successful drug discovery. In this study, we improved the tube formation assay system in three aspects. First, we used human endothelial colony forming cells (ECFCs), which are endothelial precursors that have a robust proliferative capacity and more defined angiogenic characteristics compared to mature ECs. Second, we utilized a real-time cell recorder to track the progression of tube formation for 48 hours. Third, to minimize analysis error due to the limited observation area, we used image-stitching software to increase the microscope field of view to a $2{\times}2$ stitched area from the $4{\times}$ object lens. Our advanced tube formation assay system successfully demonstrated the time-dependent dynamic progression of tube formation in the presence and absence of VEGF and FGF-2. Vatalanib, VEGF inhibitor, was tested by our assay system. Of note, $IC_{50}$ values of vatalanib was different at each observation time point. Collectively, these results indicate that our advanced tube formation assay system replicates the dynamic progression of tube formation in response to angiogenic modulators. Therefore, this new system provides a sensitive and versatile assay model for evaluating pro- or anti-angiogenic drugs.

Nondestructive Techniques for Characterization of Microstructural Evolution during Low Cycle Fatigue of Cu and Cu-Zn Alloy (Cu와 Cu-Zn 합금의 저주기피로 동안 발달한 미세조직 평가를 위한 비파괴기술)

  • Kim, Chung-Seok;Jhang, Kyung-Young;Hyun, Chang-Young
    • Journal of the Korean Society for Nondestructive Testing
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    • v.31 no.1
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    • pp.32-39
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    • 2011
  • The object of this study is to evaluate and discriminate nondestructively the dislocation substructures of Cu and Cu-Zn alloy subjected to the low-cycle-fatigue. The ultrasonic wave velocity, electrical resistivity and positron annhilation lifetime(PAL) were measured to the nondestructive testing. Cyclic fatigue test of Cu and Cu-Zn alloy with much different stacking fault energies was conducted and the correlations between dislocation behavior and nondestructive parameters were studied. Dislocation cell substructure was developed in Cu, while planar array of dislocation structure was developed in Cu-35Zn alloy only increasing dislocation density with fatigue cycles. Decrease in ultrasonic wave velocity, increase in electrical resistivity and PAL were shown because of the development of lattice defects, dislocations and vacancies, by cyclic fatigue at room temperature. In contrast to Cu-Zn alloy of the planar-array dislocation substructure showing continuous changes in the nondestructive parameters, it does not make any noticeable changes in the nondestructive parameters after the evolution of dislocation cell substructure in Cu.

International Activities for the Development of a Full Engine Simulation Program (엔진 시뮬레이션 프로그램 개발의 국제 동향)

  • Jin, Sang-Wook;Kim, Kui-Soon;Choi, Jeong-Yeol;Ahn, Iee-Ki;Yang, Soo-Seok
    • Proceedings of the Korean Society of Propulsion Engineers Conference
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    • 2007.04a
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    • pp.250-257
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    • 2007
  • The development of aircraft engine requires a lot of time and cost to estimate system attributes such as performance, reliability, stability and life. A virtual engine test based on "Numerical test cell" can extremely reduce the time and cost for the development of a hardware by coupling multidisciplinary analyses. This paper presents the development activities of full engine simulation programs in U.S.A. and Europe. NASA Glenn research center of U.S.A. leads the development efforts of NPSS(Numerical Propulsion System Simulation) by assembling the existing codes and improving their functions. VIVACE (Value Improvement through a Virtual Aeronautical Collaborative Enterprise), a consortium of universities, research centers and companies in Europe is developing the PROOSIS(PRopulsion Object Oriented SImulation Software) by integrating the various programs of the institutes. The capability for the domestic development is also estimated by surveying the current status.

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Surface Properties of Liposomes Modified with Poly(ethylenimine) (폴리에틸렌이민으로 개질된 리포솜의 표면 특성)

  • 박윤정;남다은;서동환;한희동;김태우;김문석;신병철
    • Polymer(Korea)
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    • v.28 no.6
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    • pp.502-508
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    • 2004
  • Cationic liposomes for cancer treatment have been developed in the field of chemotharpy. It was well combined on the surface of anionic tumor cell membrane by electrostatic interaction. Thus, the object of this study was to prepare the cationic liposomes capable of forming an ionic complex with the anionic cell membrane. To prepare the cationic liposomes, 1,2-distearoyl-sn-glycero-3-phosphoethanolamine (DSPE) as a cationic lipid material and polyethylenimine (PEI) as a cationic polymer were synthesized. Ionic property on the surface of liposomes was determined by the zeta potential. The adsorption characteristics of plasma protein for liposome in bovine serum were determined by the particle size and turbidity change. To estimate the stability of liposome in buffered solution, the change of particle size was measured at room temperature for seven days. The cationic liposomes were absorbed a large amount of plasma protein in bovine serum because plasma protein having anionic charge was fixed on the surface of cationic liposomes. This result indicate that the modification on the surface of liposomes using cationic polyethylenimine enhances the protein adsorption in bovine serum. Additionaly, cationic liposomes showed good stability in buffered solution for seven days.

The Effect of the Moutan Radicis Cortex on Expression of CD81 and GFAP in Injured Astrocyte (목단피(牧丹皮)가 손상된 성상신경세포의 CD81 및 GFAP의 발현에 미치는 영향)

  • Moon, Sung-Jin;Seon, Ki-Moon;Lim, Jin-Young;Song, Bong-Keun
    • The Journal of Internal Korean Medicine
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    • v.30 no.1
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    • pp.24-35
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    • 2009
  • Object : In conditions of brain infarction, irreversible axon damage occurs in the central nerve system (CNS), because gliosis makes physical and mechanical barriers. If gliosis formation could be suppressed, irreversible axon damage would be reduced. This could mean that an injured CNS could be regenerated. CD81 and GFAP have close relationships to gliosis. The increase in glial cells at CNS injury gives rise to the expression of CD81 and GFAP. CD81 was postulated to play a central role in the process of CNS scar formation. Method : In this study, the author investigated the effect of the water extract of the Moutan Radicis Cortex on regulation of CD81 and GFAP expression in injured CNS cells. MTT assay was used to examine cell viability, while RT-PCR and ELISA methods were carried out to measure the expression of CD81 and GFAP in the astrocyte. Results : We observed that water extract of the Moutan Radicis Cortex increased cell viability under hypoxia induced by $CoCl_2$ and suppressed the expression of CD81 and GFAP up-regulated by hypoxia. Conclusion : These results suggest that the Moutan Redicis Cortex could promote neural regeneration as a consequence of protecting CNS cells from hypoxia and suppressing the reactive gliosis following CNS injury.

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Immune enhancement effect of Rubus coreanus Miquel (RC) (30% EtOH extract) in RAW264.7 cells (RAW264.7 세포주를 통한 복분자 30% 에탄올 추출물의 면역기능 활성증진효과 검토)

  • Cho, Jae-Kyung;Choi, Han-Seok;Kim, Min-Su;Kim, Young-Gook;Kim, Chi-Hong;Shin, Yong-Cheol;Ko, Seong-Gyu
    • Journal of Society of Preventive Korean Medicine
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    • v.16 no.3
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    • pp.155-165
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    • 2012
  • Objective : The object of this study is to confirm the immune enhancement effect of Rubus coreanus Miquel (RC) (30% EtOH extract) on RAW264.7 mouse macrophage cell line. Methods : RAW264.7 cells were treated with $10-500{\mu}g/mL$ RC for 24 hours. Cell viability was then measured using WST assays. Levels of intracellular NO and ROS were measured by Griess reagent and DCFH-DA staining respectively. Levels of iNOS and COX-2 mRNA was determined by RT-PCR. Secretion levels of IL-$1{\beta}$ and IL-6 cytokines were evaluated by sandwich ELISA assay. Western blot analysis was performed to measure the levels of intracellular molecules related to MAPKs pathways. Results : RC suppressed the growth of RAW264.7 cells. RC increased the production of NO and ROS. RC increased the mRNA and protein levels of COX-2 and iNOS. RC augmented the levels of secreted IL-$1{\beta}$ and IL-6 cytokines. RC increased MAPKs phosphorylation. Conclusion : In summary, our result shows that RC has inflammatory effect increasing the levels of NO, ROS and secreted cytokines and activating MAPKs. Hence, RC seems to have an immune enhancement effect.

The Basis of Different Sensitivities of Ovarian Cancer Cells to the Recombinant Adenoviral Vector System Containing a Tumor-Specific L-plastin Promoter and E. coli Cytosine Deaminase Gene as a Transcription Unit

  • Chung, In-Jae
    • Biomolecules & Therapeutics
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    • v.17 no.2
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    • pp.138-143
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    • 2009
  • We have reported previously on a replication incompetent recombinant adenoviral vector, AdLPCD, in which the expression of cytosine deaminase gene (CD) is driven by the tumor-specific L-plastin promoter. AdLPCD vector had been evaluated for its efficacy of chemosensitization of ovarian cancer cells to 5-FC. In spite of the fact that ovarian cancer cells, i.e., OVCAR-3 and SK-OV-3, are capable for adenoviral transduction judged by LacZ reporter gene analysis, two cell lines demonstrated quite different sensitivities toward AdLPCD/5-FC system. In OVCAR-3 cells, infection of AdLPCD followed by exposure to 5-FC resulted in the suppression of cell growth with statistical significance. On the other hand, SK-OV-3 cells were more resistant to the CD/5-FC strategy compared with OVCAR-3 cells under the same condition. The object of study was to investigate factors that would determine the sensitivity to AdLPCD/5-FC. We evaluated conversion rate of 5-FC to 5-FU after infection of AdLPCD by HPLC analysis, $IC_{50}$ of 5-FU, the expression level of integrin receptors i.e., ${\alpha}v{\beta}3$ and ${\alpha}v{\beta}5$, and status of p53 in OVCAR-3 and SK-OV-3 cells. The results indicated that OVCAR-3 cells have few favorable features compared with SK-OV-3 cells to be more effective to the AdLPCD/5-FC strategy; higher level of ${\alpha}v{\beta}5$ integrin, higher rate of conversion of 5-FC into 5-FC, and lower $IC_{50}$ of 5-FU. The results suggest that the replacement of 5-FU with CD/5-FC in combination chemotherapy would be less toxic and much greater cytotoxicity than the conventional combination chemotherapy in some patients.