• Korean Journal of Oriental Medicine
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• v.10 no.1
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• pp.127-135
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• 2004

Hematopoietic stem cells in bone marrow form all kinds of blood cells. In traditional medicine, functions of bone marrow cells are very similar to those of Essence(精) which is a fundamental factor of physical development and reproduction. Our experiment examined the effect of deer blood on aplastic anemia induced mouse using cyclophosphamide 150 mg/kg i.p injection before experiment and then another cyclophosphamide 120 mg/kg i.p injection on day 10. Then we administrated dried deer blood in distilled water for 5 days, 9 days and 10 days. We examined blood and marrow samples. In results, deer blood showed a trend of effectiveness on recovery of red blood cells and erythropoietin although they were not statistical significant. And deer blood did not show changes in CD34.

• Korean Journal of Clinical Laboratory Science
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• v.38 no.1
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• pp.9-15
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• 2006

Peripheral bood stem cell collection (PBSCC), including peripheral blood stem cell transplantation (PBSCT), has been utilized worldwide as a very beneficial treatment method instead of allogenic Bone Marrow Transplantation (BMT) because it has many advantages such as rapid bone marrow engraftment and hematopoietic recovery, easy and safe accessibility and lower risk of rejection compared with allogenic BMT. In order to identify most the observable parameter in PBSCC, we analyzed various hematological parameters before and after PBSCC, and evaluated the correlation between the time of bone marrow engraftment and the number of CD34+ cells. Thirteen patients, who underwent 54 PBSCCs from January, 2003 to August, 2004 at Chungnam National University Hospital due to various systemic neoplasms, were analyzed in aspects of various hematological parameters including CD34+ cells using by Flow Cytometry (FCM). PBSCC harvests are described below: Mononuclear cells (MNC) $2.3{\pm}1.4{\times}10^8/kg$ and CD34+ cells $0.63{\pm}0.35{\times}10^6/kg$ on average, respectively. There was a statistical significance in Hb and Hct before and after PBSCC, but not in WBC and platelet counts. The period to reach the hematological bone marrow engraftment was 13.4(10~21) days and 19.5(11~38) days according to the criteria of absolute neutrophile counts (ANC) ${\geq}500/uL$ and platelet counts ${\geq}50,000/{\mu}L$ in peripheral blood, respectively. There was a significant correlation between the numbers of CD34+ cell and ANC (p<0.05), and a borderline significance between MNC and ANC (p=0.051). We found that a group of patients, who were infused with CD34+ cells more than $3.5{\times}10^6/kg$, reached more rapidly the period of bone marrow engraftment in platelet counts (p=0.040). This present study suggested that Hb and Hct were the most useful parameters and should be closely monitored before and after PBSCC, that a PBSCT with the dosage of more than $3.5{\times}10^6/kg$ of CD34+ cells was needed to perform successful bone marrow engraftment, and additionally that platelet counts could be more useful in indicating bone marrow engraftment than ANC.

• International Journal of Oral Biology
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• v.31 no.2
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• pp.67-72
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• 2006

Nitric oxide(NO) is a labile, uncharged, reactive radical that functions as a sensitive mediator of intercellular communication in diverse tissues. It has been reported that NO is produced by osteoblast and these results may suggest that NO is integrally involved in the regulation of osteoclast formation and osteoclast resorption activity by osteoblastic cells. We examined the effect of cytokines on NO release by mouse bone marrow cell. We also examined the effects of cytokines and sodium nitroprusside(SNP) on the formation of osteoclast-like cell from mouse bone marrow cells in culture. Cytokines stimulated NO production of mouse bone marrow cells, and N-nitro-L-arginine methyl ester, a specific inhibitor of NO synthase, suppressed the cytokine-induced NO production. SNP showed dual action in the generation of osteoclasts. The addition of $30{\mu}M$ SNP inhibited the formation of tartrate resistant acid phosphatase(TRAP)(+) multinucleated cell, whereas lower concentration($3{\mu}M$) of SNP enhanced it. Although the precise action of NO remains to be elucidated in detail, the action of NO in osteoclast generation in our studies seems to be associated, at least in part, with bone metabolism and bone pathophysiology.

• Molecules and Cells
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• v.39 no.3
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• pp.186-194
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• 2016

Epidemiological evidence suggests that bone is especially sensitive to oxidative stress, causing bone loss in the elderly. Previous studies indicated that human amnion-derived mesenchymal stem cells (HAMSCs), obtained from human amniotic membranes, exerted osteoprotective effects in vivo. However, the potential of HAMSCs as seed cells against oxidative stress-mediated dysfunction is unknown. In this study, we systemically investigated their antioxidative and osteogenic effects in vitro. Here, we demonstrated that HAMSCs significantly promoted the proliferation and osteoblastic differentiation of $H_2O_2$-induced human bone marrow mesenchymal stem cells (HBMSCs), and down-regulated the reactive oxygen species (ROS) level. Further, our results suggest that activation of the ERK1/2 MAPK signal transduction pathway is essential for both HAMSCs-mediated osteogenic and protective effects against oxidative stress-induced dysfunction in HBMSCs. U0126, a highly selective inhibitor of extracellular ERK1/2 MAPK signaling, significantly suppressed the antioxidative and osteogenic effects in HAMSCs. In conclusion, by modulating HBMSCs, HAMSCs show a strong potential in treating oxidative stress- mediated bone deficiency.

• Reproductive and Developmental Biology
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• v.35 no.1
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• pp.23-31
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• 2011

Two piglets and one juvenile pig were used to investigate closely what types of cells express green fluorescent protein (GFP) and if any, whether the GFP-tagged cells could be used for stem cell transplantation research as a middle-sized animal model in bone marrow cells of recloned GFP pigs. Bone marrow cells were recovered from the tibia, and further analyzed with various cell lineage markers to determine which cell lineage is concurrently expressing visible GFP in each individual animal. In the three animals, visible GFP were observed only in proportions of the plated cells immediately after collection, showing 41, 2 and 91% of bone marrow cells in clones #1, 2 and 3, respectively. The intensity of the visible GFP expression was variable even in an individual clone depending on cell sizes and types. The overall intensities of GFP expression were also different among the individual clones from very weak, weak to strong. Upon culture for 14 days in vitro (14DIV), some cell types showed intensive GFP expression throughout the cells; in particular, in cytoskeletons and the nucleus, on the other hand. Others are shown to be diffused GFP expression patterns only in the cytoplasm. Finally, characterization of stem cell lineage markers was carried out only in the clone #3 who showed intensive GFP expression. SSEA-1, SSEA-3, CD34, nestin and GFAP were expressed in proportions of the GFP expressing cells, but not all of them, suggesting that GFP expression occur in various cell lineages. These results indicate that targeted insertion of GFP gene should be pursued as in mouse approach to be useful for stem cell research. Furthermore, cell- or tissue-specific promoter should also be used if GFP pig is going to be meaningful for a model for stem cell transplantation.

• International Neurourology Journal
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• v.22 no.4
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• pp.260-267
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• 2018

Purpose: A major question remaining in approaches to tissue engineering and organ replacement is the role of native mobilized native cells in the regeneration process of damaged tissues and organs. The goal of this study was to compare the cell mobilizing effects of the chemokine CXCL12 and cell therapy on the urinary sphincter of nonhuman primates (NHP) with chronic intrinsic urinary sphincter dysfunction. Methods: Either autologous lenti-M-cherry labeled skeletal muscle precursor cells (skMPCs) or CXCL12 were injected directly into the sphincter complex of female NHPs with or without surgery-induced chronic urinary sphincter dysfunction (n=4/treatment condition). All monkeys had partial bone marrow transplantation with autologous lenti-green fluorescent protein (GFP) bone marrow cells prior to treatment. Labeled cells were identified, characterized and quantified using computer-assisted immunohistochemistry 6 months posttreatment. Results: GFP-labeled bone marrow cells (BMCs) were identified in the bone marrow and both BMCs and skMPCs were found in the urinary sphincter at 6-month postinjection. BMCs and skMPCs were present in the striated muscle, smooth muscle, and lamina propria/urothelium of the sphincter tissue. Sphincter injury increased the sphincter content of BMCs when analyzed 6-month postinjection. CXCL12 treatment, but not skMPCs, increased the number of BMCs in all layers of the sphincter complex (P<0.05). CXCL12 only modestly (P=0.15) increased the number of skMPCs in the sphincter complex. Conclusions: This dual labeling methodology now provides us with the tools to measure the relative number of locally injected cells versus bone marrow transplanted cells. The results of this study suggest that CXCL12 promotes mobilization of cells to the sphincter, which may contribute more to sphincter regeneration than injected cells.

• 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.

• Development and Reproduction
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• v.9 no.1
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• pp.43-48
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• 2005

Bone marrow stromal cells, a constituent of the niche for hematopoietic stem cells in bone marrow, provide various factors involved in the fate decision of the hematopoietic stem and progenitor cells. Radiation, a widely used anti-cancer therapy, provokes side effects including the damage of the blood cells. Therefore, it is necessary to recover the blood cells shortly after radiation via promoting the differentiation of hematopoietic cells. In this study, we screened genes modulated by radiation in human bone marrow stromal cells in order to understand the mechanism involved in hematopoiesis after radiation. We performed differential display method by using polymerase chain reaction(PCR) and agarose gel electrophoresis. We found plasminogen activator inhibitor-1(PAI-1) was consistently induced by radiation. The significance of the PAI-1 gene modulation is to be determined.

• Biomolecules & Therapeutics
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• v.15 no.4
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• pp.230-234
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• 2007

In anti-cancer therapies, radiotherapy and chemotherapy show a superior inhibition effect on cancer cell growth, but those are very toxic to normal tissues and organs. Particularly, drugs for neutropenia, one of chemotherapy agents, which suppress the function of bone marrow, are needed to be controlled in terms of their dosage and therapy period because of their side effect. Phellinus linteus polysaccharide (PL) has been reported to increase the number of splenocytes and bone marrow cells. PL has been shown to decrease the side effects of cyclophosphamide (CYC) treatment to the cancer patients. PL showed no effects in semisolid clonogenic assay, suggesting that PL doesn't contain substantial compounds to substitute for colony stimulating factors (CSFs). On the other hand, PL increased the expression of SCF, IL-3, GM-CSF, TPO genes. These results indicate that PL may promote the growth and proliferation of splenocytes and bone marrow cells through indirect or CSFs-dependent pathway, which may lead to a hematopoiesis.

• Journal of Gastric Cancer
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• v.6 no.4
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• pp.221-226
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• 2006

Purpose: Controversy still exists over in the prognostic significance of microscopic tumor cell dissemination in patients with cancer. This study evaluated the prognostic implication of isolated tumor cells in the bone marrow of patients with gastric cancer. Materials and Methods: Four hundred nineteen (419) patients who underwent surgery for gastric cancer between June 1998 and July 2000 were enrolled in the study. Bone marrow aspirate was obtained from the iliac crest before removal of the primary tumor. Mononuclear cells were isolated and stained with AE-1/AE-3 PAN-CYTOKERATIN. Results: Cytokeratin-positive cells were found in the bone marrow of 219 patients (52.3%). The incidence varied significantly with the depth of invasion (P=0.021) and the stage (P=0.026). The five-year survival rate of patients with cytokeratin-positive cells was 74.1% and that of patients without cytokeratin-positive cells was 81.1%(P=0.2481). There were no significant differences in the recurrence rate and the site of recurrence according to whether or not cytokeratin-positive cells were present in the bone marrow. Conclusion: The presence of cytokeratin-positive cells in the bone marrow of patients with gastric cancer did not predict outcome and recurrence. Therefore, it cannot be used as a prognostic factor.