• Journal of Veterinary Clinics
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• v.28 no.1
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• pp.57-62
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• 2011

The shortage of transplantable kidneys has many efforts to regenerate bioartificial kidneys using transgenic animals and diverse kinds of scaffolds which are important tools for cell seeding. However, there are many limitations for clinical applications so far. Recently, decellularized bioscaffolds using animal organs come into spotlight because of its many superior advantages. In current study, we produced decellularized kidney bioscaffolds of pig which is an attractive animal as a clinical model for human. We decellularized pig kidneys with 1% SDS detergent solution using peristaltic pump systems for 12h. After decellularization process, the kidney bioscaffolds preserved intact 3D morphology including glomerular structure and almost DNA from pig was entirely removed. In addition, this process could preserve micro vascular network which is necessary for cell survival. Although, additional studies for recellularization and transplantation should be required, the decellular vascularized kidney bioscaffolds might have many potentials for kidney regeneration.

• Asian Pacific Journal of Cancer Prevention
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• v.16 no.18
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• pp.8461-8465
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• 2016

Gastric cancer (GC) as the fourth most common cause of malignancies shows high rate of morbidity appropriating the second leading cause of cancer-related death worldwide. Developmental pluripotency associated-2 (DPPA2), cancer-testis antigen (CT100), is commonly expressed only in the human germ line and pluripotent embryonic cells but it is also present in a significant subset of malignant tumors. To investigate whether or not DPPA2 expression is recalled in GC, our aim in this study was to elucidate DPPA2 protein expression in gastric cancer. Fifty five GC tumor and their related margin normal tissues were recruited to evaluate DPPA2 protein expression and its probable associations with different clinicopathological features of the patients. DPPA2 was overexpressed in GC cases compared with normal tissues (P < .005). While DPPA2 expression was detected in all GC samples, its high expression was found in 23 of 55 tumor tissues (41.8%). Interestingly, 50 of 55 normal samples (90.9%) were negative for DPPA2 protein expression and remained 5 samples showed very low expression of DPPA2. DPPA2 protein expression in GC was significantly correlated with lymph node metastasis (p = 0.012). The clinical relevance of DPPA2 in GC illustrated that high level expression of this protein was associated with lymph node metastasis supporting this hypothesis that alteration in DPPA2 was associated with aggressiveness of gastric cancer and may be an early event in progression of the disease. DPPA2 may be introduced as a new marker for invasive and metastatic GCs.

• Asian Pacific Journal of Cancer Prevention
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• v.16 no.9
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• pp.3793-3797
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• 2015

Background: Chronic myeloid leukemia (CML) is a myeloproliferative disorder of pluripotent stem cells, caused by reciprocal translocation between the long arms of chromosomes 9 and 22, t(9;22)(q34;q11), known as the Philadelphia chromosome. Materials and Methods: A total of 51 CML patients were recruited in this study. Complete blood counts of all CML patients were performed to find out their total leukocytes, hemoglobin and platelets. FISH was performed for the detection of BCR-ABL fusion and cryptogenic tests using bone marrow samples were performed for the conformation of Ph (9;22)(q34;q11) and variant translocation mechanisms. Results: In cytogenetic analysis we observed that out of 51 CML patients 40 (88.9%) were Ph positive and 4 (8.88%) had Ph negative chromosomes. Mean values of WBC 134.5 $10^3/{\mu}l$, hemoglobin 10.44 mg/dl, and platelets 288.6 $10^3/{\mu}l$ were observed in this study. Conclusions: In this study, Ph positive translocation between chromosome (9:22)(q34;q11) were observed in 40 (88.9%) CML patients.

• Development and Reproduction
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• v.20 no.2
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• pp.141-147
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• 2016

Rad51 is a key component of homologous recombination (HR) to repair DNA double-strand breaks and it forms Rad51 recombinase filaments of broken single-stranded DNA to promote HR. In addition to its role in DNA repair and cell cycle progression, Rad51 contributes to the reprogramming process during the generation of induced pluripotent stem cells. In light of this, we performed reprogramming experiments to examine the effect of co-expression of Rad51 and four reprogramming factors, Oct4, Sox2, Klf4, and c-Myc, on the reprogramming efficiency. Co-expression of Rad51 significantly increased the numbers of alkaline phosphatase-positive colonies and embryonic stem cell-like colonies during the process of reprogramming. Co-expression ofRad51 significantly increased the expression of epithelial markers at an early stage of reprogramming compared with control cells. Phosphorylated histone H2AX (${\gamma}H2AX$), which initiates the DNA double-strand break repair system, was highly accumulated in reprogramming intermediates upon co-expression of Rad51. This study identified a novel role of Rad51 in enhancing the reprogramming efficiency, possibly by facilitating mesenchymal-to-epithelial transition and by regulating a DNA damage repair pathway during the early phase of the reprogramming process.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.30 no.2
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• pp.133-141
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• 2008

Stem cells have self-renewal capacity, long-term viability, and multiline age potential. Adult bone marrow contains mesenchymal stem cells. Bone marrow-derived mesenchymal stem cells (BMSCs) are progenitors of skeletal tissue components and can differentiate into adipocytes, chondrocytes, osteoblasts, and myoblasts in vitro and undergo differentiation in vivo. However, the clinical use of BMSCs has presented problems, including pain, morbidity, and low cell number upon harvest. Recent studies have identified a putative stem cell population within the adipose tissue. Human adipose tissue contains pluripotent stem cells simillar to bone marrow-derived stem cells that can differentiate toward the osteogenic, adipogenic, myogenic, and chondrogenic lineages. Human adipose tissue-derived stem cells (ATSCs) could be proposed as an alternative source of adult bone marrow stem cells, and could be obtained in large quantities, under local anesthesia, with minimal discomfort. Human adipose tissue obtained by liposuction was processed to obtain ATSCs. In this study, we compared the osteogenic differentiation of ATSCs in a specific osteogenic induction medium with that in a non-osteogenic medium. ATSCs were incubated in an osteogenic medium for 28 days to induce osteogenesis respectively. Osteogenic differentiation was assessed by von Kossa and alkaline phosphatase staining. Expression of osteocyte specific bone sialoprotein, osteocalcin, collagen type I and alkaline phosphatase, bone morphogenic protein 2, bone morphogenic protein 6 was confirmed by RT-PCR. ATSCs incubated in the osteogenic medium were stained positively for von Kossa and alkaline phosphatase staining. Expression of osteocyte specific genes was also detected. Since this cell population can be easily identified through fluorescence microscopy, it may be an ideal source of ATSCs for further experiments on stem cell biology and tissue engineering. The present results show that ADSCs have an ability to differentiate into osteoblasts. In the present study, we extend this approach to characterize adipose tissue-derived stem cells.

• Journal of Embryo Transfer
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• v.23 no.2
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• pp.67-76
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• 2008

Since the birth of Dolly using fully differentiated somatic cells as a nuclear donor, viable clones were generated successfully in many mammalian species. These achievements in animal cloning demonstrate developmental potential of terminally differentiated somatic cells. At the same time, the somatic cell nuclear transfer (SCNT) technique provides the opportunities to study basic and applied biosciences. However, the efficiency generating viable offsprings by SCNT remains extremely low. There are several explanations why cloned embryos cannot fully develop into viable animals and what factors affect developmental potency of reconstructed embryos by the SCNT technique. The most critical and persuasive explanation for inefficiency in SCNT cloning is incomplete genomic reprogramming, such as DNA methylation and histone modification. Numerous studies on genomic reprogramming demonstrated that incorrect DNA methylation and aberrant epigenetic reprogramming are considerably correlated with abnormal development of SCNT cloned embryos even though its mechanism is not fully understood. The SCNT technique is useful in cloning farm animals because pluripotent stem cells are not established in farm animal species. Therapeutic cloning combined with genetic manipulation will help to control various human diseases. Also, the SCNT technique provides a chance to overcome excessive demand for the organs by production of transgenic animals as xenotransplantation resources. Here, we describe the factors affecting the efficiency of generating cloned farm animals by the SCNT technique and discuss future directions of animal cloning by SCNT to improve the cloning efficiency.

• Applied Microscopy
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• v.33 no.2
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• pp.131-143
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• 2003

A Differentiating pathway of hemocytes in vitro and in vivo of grasshopper, Euprepocnemis shirakii was described using light and electron microscopes. In the interior of body, the stem cells of the hemopoietic organ differentiated into six types of cells respectively which are prohemoyte, plasmatocyte, granulocyte I, granulocyte II, spherulocyte and oenocytoid. The formation of these hemocytes was derived from the stem cells surrounded by a reticular cell. Hemopoietic tissue cultured in the insect media differentiated different hemocytes, but none of them underwent any mitotic division. Morphological features of the cultured cells in media were essentially the same as those of the hemocytes differentiated from the stem cells in vivo. These results were shown that each stem cell could differentiate into different types of hemocytes. It was confirmed that the stem cells possessed the pluripotent differentiation ability to directly each hemocyte, and that the once formed hemocytes in vivo and in vitro didn t undergo further transformation to other hemocytes. The maintenance of circulating hemocytes in grasshopper had been depended on the widely spreading hemopoietic organ situated in the upper surface of the dorsal alary muscle and located on the first to eighth segments.

• Molecules and Cells
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• v.19 no.1
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• pp.46-53
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• 2005

Human embryonic stem (hES) cells, unlike most cells derived from adult or fetal human tissues, represent a potentially unlimited source of various cell types for basic clinical research. To meet the increased demand for characterized hES cell lines, we established and characterized nine new lines obtained from frozen-thawed pronucleus-stage embryos. In addition, we improved the derivation efficiency from inner cell masses (to 47.4%) and optimized culture conditions for undifferentiated hES cells. After these cell lines had been maintained for over a year in vitro, they were characterized comprehensively for expression of markers of undifferentiated hES cells, karyotype, and in vitro/in vivo differentiation capacity. All of the cell lines were pluripotent, and one cell line was trisomic for chromosome 3. Improved culture techniques for hES cells should make them a good source for diverse applications in regenerative medicine, but further investigation is needed of their basic biology.

• Molecules and Cells
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• v.24 no.2
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• pp.255-260
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• 2007

Bone marrow (BM) derived mesenchymal stem cells (MSC) are pluripotent cells which can differentiate into osteogenic, adipogenic and other lineages. In spite of the broad interest, the information about the changes in BM cell composition, in particularly about the variation of MSC number and their properties in relation to the age of the donor is still controversial. The aim of this study was to investigate the age associated changes in variations of BM cell composition, phenotype and differentiation capacities of MSC using a rat model. Cell populations were characterized by flow cytometry using light scattering parameters, DNA content and a set of monoclonal antibodies. Single cell analysis was performed by conventional fluorescent microscopy. In vitro culture of MSC was established and their phenotype and capability for in vitro differentiation into osteogenic and adipogenic cells was shown. Age related changes in tibiae and femurs, amount of BM tissue, BM cell composition, proportions of separated MSC and yield of MSC in 2 weeks of in vitro culture were found. At the same time, neither change in phenotype no in differentiation capacities of MSC was registered. Age-related changes of the number of MSC should be taken into account whenever MSC are intended to be used for investigations.

• YAKHAK HOEJI
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• v.47 no.4
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• pp.234-238
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• 2003

Linarin is a main compound from Chrysanthemum zawadskii var, latilobum. However, the biological mechanisms of these activities are unclear. Because of this wide diversity of effects, it is believed that they may be exerted through pluripotent effectors of linarin. In our previous screening study, the effects of linarin on the mouse macrophages cell line, RAW 264.7 cells, were investigated. It was found that linarin could stimulate macrophages activation by the production of tumor necrosis factor (TNF). The linarin (6.25∼12.5 $\mu\textrm{g}$/mι) inhibited the production of NO in LPS-activated RAW 264.7 cells and linarin became an useful candidates for the development of new drug to treat endotoxemia and inflammation accompanied by the overproduction of NO. However, linarin-treated total lymphocyte showed cytotoxicity in a dose dependent manner between 20 $\mu\textrm{g}$/mι and 40 $\mu\textrm{g}$/mι. In this study, linarin derivative (acetylated linarin) was synthesized in order to obtain less-cytotoxicity of linarin and evaluated for their in vitro cytotoxic activity aganist mouse total lymphocyte. There was no cytotoxic activity in a dose dependent manner (20∼40 $\mu\textrm{g}$/mι) of acetylated linarin whereas linarin showed. The production of NO, however, was not the case by this modified linarin. The cell morphological change was not significantly changed in response to acetylated linarin alone and these effects were potentiated by the addition of LPS. These results suggest that acetylated linarin may be developed to be a promising new drug candidate without cytotoxicity on the basis of its activity of macrophage activation.