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A Novel Culture Model for Human Pluripotent Stem Cell Propagation on Gelatin in Placenta-conditioned Media

The propagation of human pluripotent stem cells (hPSCs) in conditioned medium derived from human cells in feeder-free culture conditions has been of ...

Graduate School of Medicine, Korea University

Frozen tumor tissues from a patient who showed rapid progression to anaplastic oligodendroglioma after near total resection of oligodendroglioma were used to examine differential expression of proteins to gain better understanding of the pathogenesis of malignant transformation.

Investigation of molecular factors associated with malignant transformation of oligodendroglioma by proteomic study of a single case of rapid tumor progression.

To investigate the underlying intratumoral diversity of molecular profiles in glioblastomas, a proteomic approach was introduced to compare samples from regions of different histological grade. Using two-dimensional gel electrophoresis (2DE) with matrix-assisted laser-desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS), we compared prospectively collected tissue samples of different histological grade areas of three glioblastoma patients. Consistent results showing relatively high expression of ubiquitin carboxyl-terminal esterase L1 in low-histological-grade areas (Grade 2 > Grades 3 and 4) and high expression of transthyretin in high-histological-grade areas (Grade 2 < Grades 3 and 4) were demonstrated. These results were confirmed by western blot (WB) analysis and immunohistochemical staining. This study provided the evidence of multifarious proteomic signatures according to regional and histological heterogeneity in glioblastomas.

Multifarious proteomic signatures and regional heterogeneity in glioblastomas.

Characterization of a rare subgroup of glioblastoma patients who survive for more than 3 years (long-term survival glioblastoma, LTSGBL, patients) may be helpful to identify prognostic factors.

Tissue expression of manganese superoxide dismutase is a candidate prognostic marker for glioblastoma.

Circulating endothelial progenitor cells (EPCs) play an important role in physiological and pathological neovascularization and may be involved in attenuating ischemic diseases. This study aimed to characterize circulating EPCs in moyamoya disease (MMD), one of the most common pediatric cerebrovascular diseases. Twenty-eight children with MMD prior to any surgical treatment and 12 healthy volunteers were recruited. Peripheral blood mononuclear cells (PBMNCs) were isolated and cultured in endothelial cell growth medium. Temporal change of phenotype of cells was analyzed on days 0 and 7. The formation of EPC clusters was evaluated on day 7. The CD34(+), CD133(+), and KDR(+) cells, and the number of EPC clusters was significantly reduced in children with MMD. In controls, CD34(+) cells were significantly decreased on day 7 compared with day 0, but in MMD they were only slightly decreased. The change in KDR(+) cells on day 7 compared with day 0 was the reverse of that for CD34(+) cells. Functional assay of EPC demonstrated less tube formation and increased senescent-like phenotype in children with MMD. Analysis of the circulating EPCs of MMD children reveals decreased level and defective function. This study suggests that circulating EPCs may be associated with MMD pathogenesis.

Decreased level and defective function of circulating endothelial progenitor cells in children with moyamoya disease.

Mel-18 is a mammalian homolog of Polycomb group (PcG) genes. Microarray analysis revealed that Mel-18 expression was induced during endothelial progenitor cell (EPC) differentiation and correlates with the expression of EC-specific protein markers. Overexpression of Mel-18 promoted EPC differentiation and angiogenic activity of ECs. Accordingly, silencing Mel-18 inhibited EC migration and tube formation in vitro. Gene expression profiling showed that Mel-18 regulates angiogenic genes including kinase insert domain receptor (KDR), claudin 5, and angiopoietin-like 2. Our findings demonstrate, for the first time, that Mel-18 plays a significant role in the angiogenic function of ECs by regulating endothelial gene expression.

Mel-18, a mammalian Polycomb gene, regulates angiogenic gene expression of endothelial cells.

Hesperidin is a flavonone glycoside found abundantly in citrus fruits that reportedly possesses anti-inflammatory, anticancer, and immune effects. Irradiation has been widely used for both diagnostic and therapeutic purposes, but it has the side effect of damaging normal cells and thereby inducing inflammation. This study was performed to investigate the effect of hesperidin on immune reactivity and nutritional status in mice with irradiation-induced inflammation. Two different concentrations (50 mg/kg and 200 mg/kg of body weight) of hesperidin were orally administered for 6 weeks to mice with or without 15-Gy irradiation treatment starting 2 weeks before irradiation. Splenocyte proliferation on Day 10 after irradiation was enhanced by supplementation with hesperidin at 50 mg/kg of body weight compared with that of the control group without irradiation on Day 30 after irradiation. The percentages of CD4(+) and CD8(+) lymphocytes in the 50 mg/kg of body weight hesperidin group tended to increase compared with the normal group. The concentration of serum cytokines (interleukin-1β, interleukin-6, and tumor necrosis factor-α) decreased in the radiation group treated with hesperidin at 50 and 200 mg/kg of body weight compared with the control group on Day 10 after irradiation. Irradiated mice fed 50 mg/kg of body weight hesperidin had significantly higher levels of total protein and albumin compared with the other groups 30 days after irradiation. In conclusion, this study suggests that hesperidin may enhance immunocompetence, have beneficial effects on nutritional status, and decrease irradiation-induced inflammation in mice.

Hesperidin partially restores impaired immune and nutritional function in irradiated mice.

Oxidative stress related to the aging process can increase the risk of degenerative disease. Red onions contain antioxidative compounds. This study was designed to investigate the effect of dietary red onion peel and/or flesh on antioxidative activity in rats. Twenty Sprague-Dawley male rats (18 weeks old) were divided into four groups. Each group was raised for 4 weeks on a red onion free control diet (ND), red onion diet containing 5% red onion peel (RP), 5% red onion flesh (RF), or 5% red onion peel+flesh (RPF). The results demonstrated that serum SOD activity was significantly increased in the RP and RPF groups, whereas glutathione peroxidase (GPx) activity was significantly higher in the RF group than in the ND group. Catalase activity and ORAC activity in liver showed upward tendency in the RP, RF, and RPF groups although the differences were not statistically significant. Liver malondialdehyde levels in the RPF group were significantly lower than those in the ND group were. In conclusion, red onion may enhance antioxidant defense mechanism through the induction of plasma SOD and GPx activities and inhibited liver lipid peroxidation. Therefore, red onion may exert important protective effects against oxidative stress related diseases.

Assessment of red onion on antioxidant activity in rat.

Radiation used in cancer treatment may cause side effects such as inflammation. Quercetin is a polyphenol that reduces inflammation. This study evaluated the recovery efficacy of quercetin on impaired immune function in irradiation-induced inflammatory mice. Quercetin administered at two concentrations of 10 and 40 mg/kg body weight was initiated 2 weeks before irradiation and was continued 30 days after irradiation. The animals exposed/not exposed to radiation were sacrificed on radiation days 10 and 30. Splenocyte proliferation, which was diminished after irradiation, was enhanced significantly by quercetin supplementation after 30 days of irradiation. Cytokine secretion increased in the radiation group compared to that in the non-radiation control group. After 30 days of radiation, interleukin (IL)-1β and IL-6 secretion decreased significantly in the radiation-quercetin groups. When quercetin was administered for 44 days, it showed a possible protective effect against irradiation-induced inflammation in mice. Quercetin could be beneficial in the recovery of irradiation-induced increases in cytokine secretion.

Effect of quercetin on impaired immune function in mice exposed to irradiation.

To evaluate the probiotic potential of Bacillus polyfermenticus CJ6 isolated from meju, a Korean traditional soybean fermentation starter, its functionality and safety were investigated. B. polyfermenticus CJ6 was sensitive to all antibiotics listed by the European Food Safety Authority. The strain was also non-hemolytic, carried no emetic toxin or enterotoxin genes, and produced no enterotoxins. The resistance of B. polyfermenticus CJ6 vegetative cells and spores to simulated gastrointestinal conditions was high (60-100% survival rate). B. polyfermenticus CJ6 produced high amounts (0.36 g as a purified lyophilized form) of gamma-polyglutamic acid (PGA). We speculate that the improved cell viability and the production of gamma-PGA have a significant correlation. Adhesion of the strain to Caco-2 and HT-29 cells was weaker than that of the reference strain (Lb. rhamnosus GG), but it was comparable to or stronger than those of reported Bacillus spp. When B. polyfermenticus CJ6 spores were given orally to mice, the number of cells excreted in the feces was 4-fold higher than the original inocula. This suggests the inoculated spores propagated within the intestinal tract of the mice. This idea was confirmed by field emission scanning electron microscopy, which revealed directly that B. polyfermenticus CJ6 cells germinated and adhered within the gastrointestinal tract of mice. Taken together, these findings suggest that B. polyfermenticus CJ6 has probiotic potential for both human consumption and use in animal feeds.

Evaluation of the probiotic potential of Bacillus polyfermenticus CJ6 isolated from Meju, a Korean soybean fermentation starter.

Non-alcoholic fatty liver disease (NAFLD) is defined as excess of fat in the liver. We investigated the effects of black soybean on the cholesterol metabolism and insulin resistance of mice fed high cholesterol/fat diets. Mice were randomly allocated into four groups that were fed different diets: the normal cholesterol/fat diet; high cholesterol/fat diets (HCD); and HCD with 1%, and 4% black soybean powder (1B-HCD, and 4B-HCD). Liver total cholesterol and triglyceride concentrations were significantly lower in the black soybean-supplemented groups than that in the HCD group. PCR revealed significantly lower hepatic SREBP2 and HMG-CoA reductase mRNA levels of black soybean-supplemented mice. Real-time PCR revealed significantly higher hepatic ABCA1 mRNA level of black soybean-supplemented mice, which may increase cholesterol efflux. Liver bile acids concentration was significantly high in the 4B-HCD group. Black soybean stimulated secretion of adiponectin, activation of pAMPK, and eliminated free fatty acids in the liver. Black soybean supplementation decreased MDA and nitrate level. The activities of SOD, catalase, and GPx were restored by black soybean supplementation. Our data strongly indicate that black soybean influences the balance between oxidative and antioxidative stress. We suggest that black soybean improves cholesterol metabolism, insulin resistance, and alleviates oxidative damage in NAFLD.

The inhibitory effect of black soybean on hepatic cholesterol accumulation in high cholesterol and high fat diet-induced non-alcoholic fatty liver disease.

Type 1 diabetes is an endocrinologic disorder characterized by uncontrolled glucose regulation and oxidative stress. Olive leaves have been studied extensively for their antioxidant activity and capacity to improve immune function. We hypothesized that olive leaf powder supplementation will be effective in inhibiting the oxidative stress and immune dysregulation in streptozotocin (STZ)-induced diabetic mice. Mice were assigned to 1 of 5 groups: control (C), STZ-induced diabetes (D), and STZ-induced diabetes supplemented with very low dose (VLOL), low dose (LOL), or high dose of olive leaf powder (HOL). Blood glucose in the VLOL and LOL groups was lower than that in the D group (P < .05). Insulin levels were increased in all experimental groups in comparison with that in the D group, (P < .05). Superoxide dismutase, glutathione peroxidase, and catalase activities were shown to decrease in the D group, whereas these were increased in the VLOL and LOL groups. Nitric oxide levels decreased in the VLOL and LOL groups, as compared with the D group. The messenger RNA expression levels of inducible nitric oxide synthase were significantly decreased in the VLOL and HOL groups, and interferon-γ levels were significantly decreased in the liver of the VLOL, LOL, and HOL groups compared with the levels in the D group. Interleukin-17 levels were significantly decreased in the VLOL and HOL groups. Th1 and Th17 cytokine levels were increased in the D group but decreased in all the experimental groups. Th2 cytokine levels were increased in all olive leaf-supplemented groups compared with those in the D group. These results indicate a reduction in the levels of proinflammatory cytokines, suggesting that olive leaves have the potential to provide therapeutic inhibition of diabetic complications.

Olive leaf down-regulates the oxidative stress and immune dysregulation in streptozotocin-induced diabetic mice.

Recent advances in dialysis and a multidisciplinary approach to pregnant patients with advanced chronic kidney disease provide a better outcome. A 38-yr-old female with autosomal dominant polycystic kidney disease (ADPKD) became pregnant. She was undergoing hemodialysis (HD) and her kidneys were massively enlarged, posing a risk of intrauterine fetal growth restriction. By means of intensive HD and optimal management of anemia, pregnancy was successfully maintained until vaginal delivery at 34.5 weeks of gestation. We discuss the special considerations involved in managing our patient with regard to the underlying ADPKD and its influence on pregnancy.

Successful pregnancy in a patient with autosomal dominant polycystic kidney disease on long-term hemodialysis.

Type 2 diabetes, characterized by hyperglycemia and hyperlipidemia, is a metabolic disease resulting from defects in both insulin secretion and insulin resistance. Recently, olive leaf has been reported as an anti-inflammatory, antioxidant, and antidiabetic agent. This study sought to investigate whether olive leaf extract can improve the insulin resistance and inflammation response in rats with type 2 diabetes induced by high-fat diet and streptozotocin. After administering olive leaf extract for 8 weeks (200 and 400 mg/kg body weight), rats given the higher dose showed significantly lower blood glucose, serum total cholesterol, and triglyceride levels compared with those of diabetic control rats (P < .05). Results of oral glucose tolerance tests, homeostasis model assessment of insulin resistance, and messenger RNA (mRNA) expression of tumor necrosis factor α and interleukin (IL) 6 in the liver show significantly decreased glucose level in rats given either dose of olive leaf extract (P < .05). Both olive leaf extract-treated groups showed significantly increased insulin receptor substrate 1 expression (P < .05). Tumor necrosis factor α, IL-6 and IL-1β mRNA expressions in epididymis adipose tissue were significantly lower in rats that received higher dose of olive leaf extract (P < .05). Lymphocyte infiltration was not observed in these rats. The results suggest that olive leaf extract may attenuate insulin resistance by suppressing mRNA expression of proinflammatory cytokines and elevating of insulin receptor substrate 1 expression.

Olive leaf extract suppresses messenger RNA expression of proinflammatory cytokines and enhances insulin receptor substrate 1 expression in the rats with streptozotocin and high-fat diet-induced diabetes.

Tuberculosis remains a serious infectious disease with primary features of pulmonary manifestation in Korea. However, duodenal tuberculosis is rare in gastrointestinal cases of extrapulmonary tuberculosis. Here, we report a case of primary duodenal tuberculosis mistaken as a malignant tumor and diagnosed with QuantiFERON-TB GOLD (Cellestis Ltd.) in an immunocompetent male patient.

A case report of primary duodenal tuberculosis mimicking a malignant tumor.

Basic fibroblast growth factor (bFGF) is a crucial factor sustaining human pluripotent stem cells (hPSCs). We designed this study to search the substitutive factors other than bFGF for the maintenance of hPSCs by using human placenta-derived conditioned medium without exogenous bFGF (hPCCM-), containing chemokine (C-X-C motif) receptor 2 (CXCR2) ligands, including interleukin (IL)-8 and growth-related oncogene α (GROα), which were developed on the basis of our previous studies. First, we confirmed that IL-8 and/or GROα play independent roles to preserve the phenotype of hPSCs. Then, we tried CXCR2 blockage of hPSCs in hPCCM- and verified the significant decrease of pluripotency-associated genes expression and the proliferation of hPSCs. Interestingly, CXCR2 suppression of hPSCs in mTeSR™1 containing exogenous bFGF decreased the proliferation of hPSCs while maintaining pluripotency characteristics. Lastly, we found that hPSCs proliferated robustly for more than 35 passages in hPCCM- on a gelatin substratum. Higher CXCR2 expression of hPSCs cultured in hPCCM- than those in mTeSR™1 was observable. Our findings suggest that CXCR2 and its related ligands might be novel factors comparable to bFGF supporting the characteristics of hPSCs and hPCCM- might be useful for the maintenance of hPSCs as well as for the accurate evaluation of CXCR2 role in hPSCs without the confounding influence of exogenous bFGF.

Ji-Hye Jung

Department of Biomedical Science

Graduate School of Medicine, Korea University

A Novel Culture Model for Human Pluripotent Stem Cell Propagation on Gelatin in Placenta-conditioned Media

Ji-Hye Jung

.css-o250i3{font-size:18px;font-family:Open Sans,sans-serif;line-height:21.6px;}Department of Biomedical Science

Graduate School of Medicine, Korea University

A Novel Culture Model for Human Pluripotent Stem Cell Propagation on Gelatin in Placenta-conditioned Media

Department of Biomedical Science