• Title/Summary/Keyword: Ex vivo

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Proliferation, Apoptosis, and Telomerase Activity in Human Cord Blood CD34+ Cells Cultured with Combinations of Various Cytokines

  • Ahn, Myung-Ju;Lee, Hye-Sook;Jang, Mi-Yune;Choi, Jung-Hye;Lee, Young-Yeul;Park, Hyung-Bae;Lee, Yong-Sung
    • Journal of Microbiology and Biotechnology
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    • v.13 no.3
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    • pp.422-428
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    • 2003
  • Umbilical cord blood (UCB), a rich source of hematopoietic stem/progenitor cells, has been proposed as an alternative to bone marrow and peripheral blood for transplantation treatment. Ex vivo expansion of cord blood stem cells could make the use of cord blood transplant feasible even for adult patients. However, the optimal cytokine cocktail for expansion of stem cells is yet to be established. This study compares proliferation, apoptosis, and telomerase activities in human cord blood stem cells cultured ex vivo with FLT3 ligand (FL)/thrombopoietin (TPO) or FL/TPO/stem cell factor (SCF), with a view to determine optimal combination of cytokines. CD34+ cells were cultured in DMEM containing either FL (50 ng/ml) and TPO (10 ng/ml) (FT group) or FL (50 ng/ml), TPO (10 ng/ml) and SCF (50 ng/ml) (FTS group). The cell proliferation rate was ten times higher in the FTS group. Although cells cultured with the two different combinations of cytokines were maintained for a long term (up to 8 weeks), a large number of cells underwent differentiation during this period. Cells cultured in FTS displayed lower levels of apoptosis compared to those of the FT group during the Initial 7 days of culture. The CD34+ fraction in both groups was markedly decreased to $21-30\%$ , and only $5-6\%$ was detected at 14 days of culture. Telomerase activity detected in human CD34+ cord blood at low levels was upregulated during the early phase of culture and decreased to baseline levels in the later phase. The telomerase activity of cord blood cultured in FT was lower than that of the FTS group. Our results suggest that, on adding stem cell factors to the FT cytokines, cultured CD34+ cord blood cells display a greater degree of cell proliferation and decreased apoptosis. However, during CD34+ cord blood cell culture, a Barge number of cells undergo differentiation, indicating that more potent novel cytokines or new culture conditioning methods should be developed to maintain their ability to engraft and sustain long-term hematopoiesis.

Cryopreservation of in vitro matured oocytes after ex vivo oocyte retrieval from gynecologic cancer patients undergoing radical surgery

  • Park, Chan Woo;Lee, Sun Hee;Yang, Kwang Moon;Lee, In Ho;Lim, Kyung Teak;Lee, Ki Heon;Kim, Tae Jin
    • Clinical and Experimental Reproductive Medicine
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    • v.43 no.2
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    • pp.119-125
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    • 2016
  • Objective: The aim of this study was to report a case series of in vitro matured (IVM) oocyte freezing in gynecologic cancer patients undergoing radical surgery under time constraints as an option for fertility preservation (FP). Methods: Case series report. University-based in vitro fertilization center. Six gynecologic cancer patients who were scheduled to undergo radical surgery the next day were referred for FP. The patients had endometrial (n=2), ovarian (n=3), and double primary endometrial and ovarian (n=1) cancer. Ex vivo retrieval of immature oocytes from macroscopically normal ovarian tissue was followed by mature oocyte freezing after IVM or embryo freezing with intracytoplasmic sperm injection. Results: A total of 53 oocytes were retrieved from five patients, with a mean of 10.6 oocytes per patient. After IVM, a total of 36 mature oocytes were obtained, demonstrating a 67.9% maturation rate. With regard to the ovarian cancer patients, seven IVM oocytes were frozen from patient 3, who had stage IC cancer, whereas one IVM oocyte was frozen from patient 4, who had stage IV cancer despite being of a similar age. With regard to the endometrial cancer patients, 15 IVM oocytes from patient 1 were frozen. Five embryos were frozen after the fertilization of IVM oocytes from patient 6. Conclusion: Immature oocytes can be successfully retrieved ex vivo from macroscopically normal ovarian tissue before radical surgery. IVM oocyte freezing provides a possible FP option in patients with advanced-stage endometrial or ovarian cancer without the risk of cancer cell spillage or time delays.

Direction of Tissue Contraction after Microwave Ablation: A Comparative Experimental Study in Ex Vivo Bovine Liver

  • Junhyok Lee;Hyunchul Rhim;Min Woo Lee;Tae Wook Kang;Kyoung Doo Song;Jeong Kyong Lee
    • Korean Journal of Radiology
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    • v.23 no.1
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    • pp.42-51
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    • 2022
  • Objective: This study aimed to investigate the direction of tissue contraction after microwave ablation in ex vivo bovine liver models. Materials and Methods: Ablation procedures were conducted in a total of 90 sites in ex vivo bovine liver models, including the surface (n = 60) and parenchyma (n = 30), to examine the direction of contraction of the tissue in the peripheral and central regions from the microwave antenna. Three commercially available 2.45-GHz microwave systems (Emprint, Neuwave, and Surblate) were used. For surface ablation, the lengths of two overlapped square markers were measured after 2.5- and 5-minutes ablations (n = 10 ablations for each system for each ablation time). For parenchyma ablation, seven predetermined distances between the markers were measured on the cutting plane after 5- and 10-minutes ablations (n = 5 ablations for each system for each ablation time). The contraction in the radial and longitudinal directions and the sphericity index (SI) of the ablation zones were compared between the three systems using analysis of variance. Results: In the surface ablation experiment, the mean longitudinal contraction ratio and SI from a 5-minutes ablation using the Emprint, Neuwave, and Surblate systems were 28.92% and 1.04, 20.10% and 0.53, and 24.90% and 0.45, respectively (p < 0.001). A positive correlation between longitudinal contraction and SI was noted, and a similar radial contraction was observed. In the parenchyma ablation experiment, the mean longitudinal contraction ratio and SI from a 10-minutes ablation using the three pieces of equipment were 38.60% and 1.06, 32.45% and 0.61, and 28.50% and 0.50, respectively (p < 0.001). There was a significant difference in the longitudinal contraction properties, whereas there was no significant difference in the radial contraction properties. Conclusion: The degree of longitudinal contraction showed significant differences depending on the microwave ablation equipment, which may affect the SI of the ablation zone.

Adipose Tissue-Derived Mesenchymal Stromal Cells from Ex-Morbidly Obese Individuals Instruct Macrophages towards a M2-Like Profile In Vitro

  • Daiana V. Lopes Alves;Cesar Claudio-da-Silva;Marcelo C. A. Souza;Rosa T. Pinho;Wellington Seguins da Silva;Periela S. Sousa-Vasconcelos;Radovan Borojevic;Carmen M. Nogueira;Helio dos S. Dutra;Christina M. Takiya;Danielle C. Bonfim;Maria Isabel D. Rossi
    • International Journal of Stem Cells
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    • v.16 no.4
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    • pp.425-437
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    • 2023
  • Obesity, which continues to increase worldwide, was shown to irreversibly impair the differentiation potential and angiogenic properties of adipose tissue mesenchymal stromal cells (ADSCs). Because these cells are intended for regenerative medicine, especially for the treatment of inflammatory conditions, and the effects of obesity on the immunomodulatory properties of ADSCs are not yet clear, here we investigated how ADSCs isolated from former obese subjects (Ex-Ob) would influence macrophage differentiation and polarization, since these cells are the main instructors of inflammatory responses. Analysis of the subcutaneous adipose tissue (SAT) of overweight (OW) and Ex-Ob subjects showed the maintenance of approximately twice as many macrophages in Ex-Ob SAT, contained within the CD68+/FXIII-A- inflammatory pool. Despite it, in vitro, coculture experiments revealed that Ex-Ob ADSCs instructed monocyte differentiation into a M2-like profile, and under inflammatory conditions induced by LPS treatment, inhibited HLA-DR upregulation by resting M0 macrophages, originated a similar percentage of TNF-α+ cells, and inhibited IL-10 secretion, similar to OW-ADSCs and BMSCs, which were used for comparison, as these are the main alternative cell types available for therapeutic purposes. Our results showed that Ex-Ob ADSCs mirrored OW-ADSCs in macrophage education, favoring the M2 immunophenotype and a mixed (M1/M2) secretory response. These results have translational potential, since they provide evidence that ADSCs from both Ex-Ob and OW subjects can be used in regenerative medicine in eligible therapies. Further in vivo studies will be fundamental to validate these observations.

Numerical Modeling of Compression-Controlled Low-level Laser Probe for Increasing Photon Density in Soft Tissue

  • Kwon, Ki-Woon;Son, Tae-Yoon;Yeo, Chang-Min;Jung, Byung-Jo
    • Journal of the Optical Society of Korea
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    • v.15 no.4
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    • pp.321-328
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    • 2011
  • Various methods have been investigated to increase photon density in soft tissue, an important factor in low-level laser therapy. Previously we developed a compression-controlled low-level laser probe (CCLLP) utilizing mechanical negative compression, and experimentally verified its efficacy. In this study, we used Bezier curves to numerically simulate the skin deformation and photon density variation generated by the CCLLP. In addition, we numerically modeled changes in optical coefficients due to skin deformation using a linearization technique with appropriate parameterization. The simulated results were consistent with both human in vivo and porcine ex vivo experimental results, confirming the efficacy of the CCLLP.

Electrochemical Diagnosis of Magnesium Ion in Fish Liver and Mice Droppings

  • Lee, Chang-Hyun;Ly, Suw Young
    • Journal of the Korean Applied Science and Technology
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    • v.31 no.4
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    • pp.663-668
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    • 2014
  • An in-vivo diagnosis of trace Mg(II) ion was performed using a low-cost and environment-friendly voltammetric method, using a graphite counter and reference electrodes and a fluorine-immobilized graphite working electrode, and clean deep seawater was used as an electrolyte solution. Under optimum conditions, the analytical working ranges attained microgram ranges, and a detection limit of $80.6ugL^{-1}$ was obtained using stripping voltammety with 60 sec accumulation time. Ex-vivo application was performed on fish liver and mice droppings. The developed techniques can be applicable to tumor cell analysis.

Anti-platelet Effects of Mixtures of Onion and Aloe Extract (양파와 알로에 복합물이 혈소판 응집에 미치는 영향)

  • Lee, Bo Kyung;Lee, Do-Seop;Ha, Soyoung;Park, Se Won;Jung, Yi-Sook
    • YAKHAK HOEJI
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    • v.58 no.5
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    • pp.322-327
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    • 2014
  • In this study, we investigated whether the mixtures of onion water extract and aloe ethanol extracts have antiplatelet activities. The mixtures inhibited collagen- and thrombin-induced rat platelet aggregation in vitro. Additionally, the oral administration of the mixtures inhibited platelet aggregation induced by collagen ex vivo but not prolonged mouse tail vein bleeding time in vivo. These results suggest that the combination of onion and aloe extracts has a potential to be a preventive agent against platelet-mediated disorders.

Synthesis of dimeric fluorescent TSPO ligand for detection of glioma

  • Tien Tan Bui;Hee-Kwon Kim
    • Journal of Radiopharmaceuticals and Molecular Probes
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    • v.7 no.1
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    • pp.56-65
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    • 2021
  • TSPO, an 18-kDa translocator protein, is a peripheral-type benzodiazepine receptor that has been associated to a variety of biological activities such as apoptosis, steroidogenesis, and cell proliferation. Because TSPO overexpression has been found in various forms of cancer, it has recently become one of the most appealing biological targets for cancer therapies and detection. In order to create new optical imaging agents for improved diagnostics, we synthesized a novel dimeric fluorescent TSPO ligand based on PRB28 structure and SCy5.5. Following the preparation of the novel TSPO ligand, in vivo and ex vivo imaging tests were performed to examine the tumor uptake characteristics of the fluorescent TSPO ligand in a glioma animal model, and it was found that novel TSPO ligand was accumulated in glioma. These results suggested that novel dimeric fluorescent TSPO ligand will be applied to detect glioma.

Use of Cell-Penetrating Peptides in Dendritic Cell-Based Vaccination

  • Sangho Lim;Ja-Hyun Koo;Je-Min Choi
    • IMMUNE NETWORK
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    • v.16 no.1
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    • pp.33-43
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    • 2016
  • Cell-penetrating peptides (CPPs) are short amino acids that have been widely used to deliver macromolecules such as proteins, peptides, DNA, or RNA, to control cellular behavior for therapeutic purposes. CPPs have been used to treat immunological diseases through the delivery of immune modulatory molecules in vivo. Their intracellular delivery efficiency is highly synergistic with the cellular characteristics of the dendritic cells (DCs), which actively uptake foreign antigens. DC-based vaccines are primarily generated by pulsing DCs ex vivo with various immunomodulatory antigens. CPP conjugation to antigens would increase DC uptake as well as antigen processing and presentation on both MHC class II and MHC class I molecules, leading to antigen specific CD4+ and CD8+ T cell responses. CPP-antigen based DC vaccination is considered a promising tool for cancer immunotherapy due to the enhanced CTL response. In this review, we discuss the various applications of CPPs in immune modulation and DC vaccination, and highlight the advantages and limitations of the current CPP-based DC vaccination.

Novel Antioxidants and Atherosclerosis

  • Parthasarathy Sampath;Santanam Nalini
    • Proceedings of the Korean Society of Food Science and Nutrition Conference
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    • 2004.11a
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    • pp.265-270
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    • 2004
  • Coronary heart disease (CHD) has been the number one killer in western society for a long time, and CHD in most instances is due to atherosclerosis. One of the earliest events in atherogenesis is the intracellular accumulation of lipids, particularly cholesterol esters, in the aortic intima. The lipids presumably came from the uptake of plasma lipoproteins, particularly from LDL. These foam cells were identified as being predominantly as macrophages. Currently, it is believed that oxidation of low density lipoprotein (LDL) might contribute to the generation of foam cells. An outcome of the oxidation hypothesis is that the consumption of antioxidants would be beneficial. In this study, Boldine, an alkaloid of Peumus boldus was tested for their antioxidant potency both in, in vitro oxidation system and in mouse models. Boldine decreased the ex-vivo oxidation of Low-density lipoprotein (LDL). In vivo studies were performed to study the effect of these compounds on the atherosclerotic lesion formation in LDL r-/- mice. Three groups of LDL r-/- mice (N=12 each) were fed an atherogenic diet. Group 1 was given vehicle and group 2 and 3 were given 1 and 5 mg of Boldine/day in addition to the atherogenic diet. The results indicated that there was a decrease in lesion formation reaching a 40% reduction due to Boldine compared to controls. The in vivo tolerance of Boldine in humans (has been used as an herbal medicine in other diseases) should make it an attractive alternative to vitamin E.

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