• Development and Reproduction
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• v.15 no.4
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• pp.339-347
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• 2011

Human eyelid adipose-derived stem cells (hEAs) and amniotic mesenchymal stem cells (hAMs) are very valuable sources for the cell therapeutics. Both types of cells have a great proliferating ability in vitro and a multipotency to differentiate into adipocytes, osteoblasts and chondrocytes. In the present study, we evaluated their stem cell characteristics after long-time cryopreservation for 6, 12 and 24 months. When frozen-thawed cells were cultivated in vitro, their cumulative cell number and doubling time were similar to freshly prepared cells. Also they expressed stem cell-related genes of SCF, NANOG, OCT4, and TERT, ectoderm-related genes of NCAM and FGF5, mesoderm/endoderm-related genes of CK18 and VIM, and immune-related genes of HLA-ABC and ${\beta}$2M. Following differentiation culture in appropriate culture media for 2-3 weeks, both types of cells exhibited well differentiation into adipocyte, osteoblast, and chondrocyte, as revealed by adipogenic, osteogenic or chondrogenic-specific staining and related genes, respectively. In conclusion, even after long-term storage hEAs and hAMs could maintain their stem cell characteristics, suggesting that they might be suitable for clinical application based on stem cell therapy.

• Journal of Nutrition and Health
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• v.39 no.8
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• pp.733-741
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• 2006

The study was conducted to investigate the effects of dietary calcium and soy isoflavone on body fat and lipid metabolism in high fat-induced obesity. Four week old female C57/BL6J mice, known as a good model of diet-induced obesity, were fed low Ca and high fat diet for 6 weeks. After induced obesity, mice were divided into six groups according to diets varying calcium contents (0.1 or 1.5%) and genistein contents (0 or 500 or 1,000 ppm). Body weight, fat pad (perirenal fat and parameterial fat), adipocyte size, serum total lipid and total cholesterol were significantly decreased by both high Ca intake and genistein supplementation. However, the effect of genistein supplementation showed in low Ca-fed groups. Serum LDL-cholesterol and TG were significantly decreased by high Ca intake and genistein supplementation, respectively. In liver, lipogenic enzymes (fatty acid synthase and malic enzyme) activity and TG were significantly decreased by both high Ca intake and genistein supplementation. This inhibitory effect of genistein on lipogenic enzymes showed in low Ca-fed groups. But liver total cholesterol and total lipid were significantly decreased by high Ca intake and genistein supplementation, respectively. Fecal excretion of total lipid, total cholesterol and TG were significantly increased by high Ca intake, not by genistein supplementation. In conclusion, high calcium intake and genistein supplement may be beneficial for suppression of obesity through direct anti-adipogenesis by decreasing fat weight and size and indirect anti-lipo-genesis by inhibiting lipogenic enzymes activity and improving lipid profile.

• Biomolecules & Therapeutics
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• v.18 no.1
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• pp.56-64
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• 2010

Pentoxifylline (PTX) has been used for the local reduction of fat tissue in the clinical setting. However, its safety and efficacy have not been proven. The aim of this study was to evaluate the effects of PTX on cell lines established from fat tissue. Newly cultured human preadipocytes and adipocytes from subcutaneous abdominal fat in addition to purchased human lung fibroblasts and keratinocytes were treated with PTX at different concentrations. Cell viability was determined using the Cell counting kit (CCK)-8 assay and lipolysis was evaluated using an Elisa kit. DNA fragmentation, Western blot analysis, Hoechst and Propidium Iodide (PI) staining and fluorescence activated cell scanning analysis were performed to confirm apoptosis. The viability of adipocytes, preadipocytes, keratinocytes and fibroblasts was markedly decreased at concentrations of PTX above 20 mM. Apoptosis was induced at concentrations of PTX over 40 mM in all cell lines. Lipolysis was increased by 60% at concentrations of PTX of 20 mM compared to the control. In conclusion, the results of this study showed that 20 mM of PTX induced lipolysis. At concentrations over 20 mM, PTX reduced the viability of all cells studied including: adipocytes, preadipocytes, fibroblasts and keratinocytes, in a non-specific manner.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.32 no.4
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• pp.327-333
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• 2006

Future cell-based therapies such as tissue engineering will benefit from a source of autogenous pluripotent stem cells. There are embryonic stem cells (ESC) and autologous adult stem cells, two general types of stem cells potentilally useful for these applications. But practical use of ESC is limited due to potential problems of cell regulation and ethical considerations. To get bone marrow stem cells is relatively burden to patients because of pain, anesthesia requirement. The ideal stem cells are required of such as the following advantages: easy to obtain, minimal patient discomfort and a capability of yielding enough cell numbers. Adipose autologus tissue taken from intraoral fatty pad or abdomen may represent such a source. Our study designed to demonstrate the ability of human adipose tissue-derived stromal cells (hATSC) from human abdominal adipose tissue diffentiating into osteocyte and adipocyte under culture in vitro conditions. As a result of experiment, we identified stromal cell derived adipose tissue has the multilineage potentiality under appropriate culture conditions. And the adipose stromal cells expressed several mesenchymal stem cell related antigen (CD29, CD44) reactions. Secondary, we compared the culture results of a group of hATSC stimulated with TGF-${\beta}$1, bFGF with a hATSC group without growth factors to confirm whether cytokines have a important role of the proliferation in osteogenic differentiation. The role of cytokines such as TGF-${\beta}$1, bFGF increased hATSC's osteogenic differentiation especially when TGF-${\beta}$1 and bFGF were used together. These results suggest that adipose stromal cells with growth factors could be efficiently available for cell-based bone regeneration.

• IMMUNE NETWORK
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• v.16 no.3
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• pp.189-194
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• 2016

Obesity is characterized as an accumulation of adipose tissue mass represented by chronic, low-grade inflammation. Obesity-derived inflammation involves chemokines as important regulators contributing to the pathophysiology of obesity-related diseases such as cardiovascular disease, diabetes and some cancers. The obesity-driven chemokine network is poorly understood. Here, we identified the profiles of chemokine signature between human preadipocytes and adipocytes, using PCR arrays and qRT-PCR. Both preadipocytes and adipocytes showed absent or low levels in chemokine receptors in spite of some changes. On the other hand, the chemokine levels of CCL2, CCL7-8, CCL11, CXCL1-3, CXCL6 and CXCL10-11 were dominantly expressed in preadipocytes compared to adipocytes. Interestingly, CXCL14 was the most dominant chemokine expressed in adipocytes compared to preadipocytes. Moreover, there is significantly higher protein level of CXCL14 in conditioned media from adipocytes. In addition, we analyzed the data of the chemokine signatures in adipocytes obtained from healthy lean and obese postmenopausal women based on Gene Expression Omnibus (GEO) dataset. Adipocytes from obese individuals had significantly higher levels in chemokine signature as follows: CCL2, CCL13, CCL18-19, CCL23, CCL26, CXCL1, CXCL3 and CXCL14, as compared to those from lean ones. Also, among the chemokine networks, CXCL14 appeared to be the highest levels in adipocytes from both lean and obese women. Taken together, these results identify CXCL14 as an important chemokine induced during adipogenesis, requiring further research elucidating its potential therapeutic benefits in obesity.

• Journal of Life Science
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• v.21 no.9
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• pp.1346-1353
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• 2011

Sophora tonkinensis Gapnep has been used as a traditional herbal medicine in oriental regions since ancient times. In this study, the effect and mechanism of the MeOH extract of Sophora tonkinensis Gapnep (STME) on adipocite differentiation and adipogenesis in 3T3-L1 preadipocites were investigated. Treatment with STME in the concentration range of 0-200 ${\mu}g$/ml significantly inhibited the differentiation of 3T3-L1 preadipocites in a dose-dependent manner, as determined by a decrease in intracellular lipid droplets and lipid contents measured by Oil Red O staining. In association with the inhibitory effect of lipid accumulation, the expressions of the proteins concerned with adipogenesis in 3T3-L1 preadipocites were also investigated. Treatment with STME reduced the expressions of peroxisome proliferator-activated receptor ${\gamma}$ (PPAR${\gamma}$), cytidine-cytidine-adenosine-adenosine-thymine (CCAAT)/enhancer-binding proteins ${\alpha}$ and ${\beta}$ (C/EBP${\alpha}$ and C/EBP${\beta}$) and sterol regulatory element binding protein (SREBP), which are adipocyte specific markers. In flow cytometry analysis, the inhibitory effect of differentiation was caused by G1 arrest and following mitotic clonal expansion cease. Therefore, we also investigated the alteration of G1 phase arrest-related proteins. As a result, the expression of p21 protein was significantly increased, while the expressions of Cdk2, E2F-1 and phospho-Rb were reduced in a dose-dependent manner in STME treated 3T3-L1 cells. According to these results, STME might inhibit differentiation through G1 arrest in 3T3-L1 preadipocytes adipogenesis, and further studies, which are in progress, have to be completed to identify the active compounds.

• Biomedical Science Letters
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• v.15 no.3
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• pp.171-177
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• 2009

This study investigated whether increased adiposity is prevented by estrogen replacement in female ovariectomized (OVX) C57BL/6J mice, an animal model of human menopause and whether these metabolic changes reflect the inhibitory action of estrogen on peroxisome proliferator-activated receptor $\gamma$ ($PPAR{\gamma}$)-regulated gene expression. Treatment of $17{\beta}$-estradiol for the last one week of the experiment decreased high fat diet-induced body weight gain and white adipose tissue mass compared to OVX control mice. Histological analysis showed that administration of $17{\beta}$-estradiol to mice decreased the size of adipocytes in parametrial adipose tissue versus OVX control mice. In addition, $17{\beta}$-estradiol reduced the adipose expression of $PPAR{\gamma}$ as well as $PPAR{\gamma}$ target genes such as adipocyte fatty acid binding protein and tumor necrosis factor $\alpha$. These results suggest that $17{\beta}$-estradiol may inhibit adiposity through reducing the $PPAR{\gamma}$ activities in female OVX mice.

• Biomedical Science Letters
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• v.11 no.2
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• pp.89-101
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• 2005

Obesity is increasing worldwide and has become a major health burden in Western societies affecting every third American and every fifth European. Obesity makes a major contribution to morbidity and mortality, predisposing individuals to cardiovascular disease and diabetes. Many new substances are currently being investigated for their usefulness in the pharmacotherapy of obesity. Most anti-obesity drugs can be divided into four groups: those that reduce food intake; those that alter metabolism; those that increase thermogenesis; and those that regulate hormone involved in feeding behavior. In this article we review these and other agents available in various countries for the treatment of obesity. Perhaps more importantly, we have focussed on areas of potential productivity in the future. Over the last 5 or so years, this impetus in obesity research has provided us with exciting new drugs targets involved in the regulation of feeding behavior and cellular mechanism involved in energy expenditure. Recent development in the quest for control of human obesity include the discovery of hormones, neuropeptides, receptors and transcription factors involved in feeding behavior, metabolic rate and adipocyte development. For developing new, perhaps even more specific pharmacological agents, further research is needed to understand the individual different genetic and physiological basis of obesity. It remains the hope of research scientists that in the not too distant future we shall see a new class of anti-obesity drugs arising logically from the molecular biology revolutions.

• CELLMED
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• v.8 no.3
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• pp.11.1-11.6
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• 2018

In vitro anti-obesity effects of anti-cancer (AC) functional kimchi in differentiated 3T3-L1 adipocytes were studied. We constructed three experimental groups: Control, standardized kimchi (SK), and AC functional kimchi (A-FK) that included active ingredients and Lactobacillus plantarum. Kimchi extracts did not show any cytotoxicity in pre-adipocytes in the concentration range of 1 - 5 mg/mL. A-FK significantly reduced fat droplet formation and absorbance in differentiated 3T3-L1 adipocytes, as shown by Oil red O staining, compared to Control and SK (P < 0.05). SK and A-FK reduced adipo-/lipogenesis related genes such as $C/EBP{\alpha}$, SREBP-1, LPL, and $LXR{\alpha}$ compared to Control (P < 0.05). Especially, A-FK more greatly reduced SREBP-1 and LPL compared to SK (P < 0.05). A-FK up-regulated the ${\beta}$-oxidation related gene CPT-1c and down-regulated the pro-inflammatory cytokine IL-6 compared to Control (P < 0.05). Based on the results, A-FK exhibited anti-obesity effects by inhibiting fat droplet formation and adipo-/lipogenesis related genes by regulating the ${\beta}$-oxidation related gene CPT-1c and pro-inflammatory cytokine IL-6. In previous studies, A-FK kimchi already exhibited a strong anti-cancer effect. These results indicate that A-FK increased anti-obesity activity in this model system due to its functional ingredients and anti-cancer functionality.

• Asian-Australasian Journal of Animal Sciences
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• v.20 no.6
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• pp.850-855
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• 2007

Leptin is the adipocyte-specific product of the obese gene and plays a major role in food intake and energy metabolism. Leptin research was mainly focused on mammalian species, but understanding of leptin and its function in poultry is very poor. In this study, the duck leptin gene was amplified using the reverse transcription-polymerase chain reaction (RT-PCR) from duck liver RNA. The cDNA fragment was inserted into the pET-28a expression vector, and the resulting plasmid was expressed in Escherichia coli BL21 (DE3). Experimental mice were given an intraperitoneal injection of 10 mg/kg leptin dissolved in phosphate buffered saline (PBS), while the control mice were injected with PBS. The effect of leptin on food intake, body weight and fatty deposition in mice was detected. Sequence analysis revealed that duck leptin had a length of 438 nucleotides which encoded a peptide with 146 amino acid residues. The sequence shares highly homology to other animals. The coding sequence of duck leptin was 84 and 86% identical to human and pig leptin nucleotides sequence. Highest identity was with the rat coding sequence (95%). The identity of the amino acid sequence was 84, 82 and 96% respectively compared to that of the human, pig and rat. Results of SDS-PAGE analysis indicated that a fusion protein was specifically expressed in E. coli BL21 (DE3). The purified product was found to be biologically active during tests. Continuous administration of recombinant duck leptin inhibited food intake. Despite the decrease of food intake, leptin significantly induced body weight and fatty deposition. These changes were accompanied by a significant down-secretion of plasma glucose, cholesterol, triglyceride and insulin levels in mice. The observations provide evidence for an inhibitory effect of leptin in the regulation of food intake and for a potential role of duck leptin in the regulation of lipogenesis.