This study examines the effects on fertilization rate (FR), hatching rate (HR), and normal individual rate after artificial fertilization using frozen thawed sperm according to the cryoprotectant (DMSO) concentration and the period of cryopreserved sperm of longtooth grouper, Epinephelus bruneus. Performing artificial fertilization using frozen-thawed sperm, after freezing the sperm at different DMSO concentration of 5.0%, 7.5%, 10.0% respectively, FR were (DMSO 5.0%: 99.5±0.8%, DMSO 7.5%: 99.5±0.7%, and DMSO 10.0%: 99.6±0.6%). The results are not significantly different from the control fresh sperm (100%). HR also (DMSO 5.0%: 96.2±2.3%, DMSO 7.5%: 95.3±3.6%, 10.0%: 96.6±1.8%) were not significantly different in each group. The normal individual rate after hatching using with control fresh sperm (98.4%±0.5) and DMSO concentration level of 5.0% (97.8±0.1%) were not significantly different. However, with 7.5% (97.2±0.6%) and 10.0% DMSO concentrations (95.9±0.2%) are lower than the normal individual rate after hatching observed in the control and 5.0% DMSO. Performing artificial fertilization using frozen-thawed sperm at different frozen period (2 days, 2 years, and 3 years), 10% DMSO FR and HR of 3 years (FR; 66.8±1.8%, HR: 82.0±12.9%) and 2 years (FR; 78.5±14.8%, HR: 79.3±0.6%) cryopreserved sperm were lower than control (FR; 100%, HR: 91.1±3.6%) and 2 days cryopreserved sperm (FR; 99.6±0.6%, HR: 96.6±1.8%). These results suggest suitable DMSO concentration ranges of cryopreservation sperm for E. bruneus is 5 to 10% and with 2 to 3 years cryopreservation period, cryopreservation sperm can be useful for seed production.

Korea Ministry of Environment (Korea MOE) enacted the “Greenhouse Gas and Energy Target Management System (GETMS), which requires annual GHG reporting to establish GHG reduction targets for large-scale business places (458) emitting large amount of greenhouse gases (60% of total amount in Korea). The waste sector has higher potential for reduction of greenhouse gases compared to other sectors. Thus, this paper reviewed the methodologies modified based on national guidelines and estimated the greenhouse gas emissions for three categories of the waste sector in Daejeon Metropolitan City (DMC), South Korea. Further analysis for basic unit, i.e., greenhouse gas emissions per ton of solid waste, wastewater, and purified water in the waste sector was conducted to figure out main contributors for GHG emissions. Direct emissions (Scope 1) and indirect emissions (Scope 2) of 11 environmental infrastructures managed by DMC were selected for quantifying and managing of regional GHG emissions. The annual estimation for greenhouse gas emissions in the waste sector in DMC with a population of 1.52 million people was 254,235 tons CO₂ equiv. per year, which includes the main contributor of wastewater treatment 78,063 tons CO2 equiv., waste incineration 76,186 tons CO₂ equiv., and managed waste disposal sites 70,455 tons CO₂ equiv. Basic unit showed that most contributors were waste incineration, followed by the waste disposal site, biological treatment of solid waste, wastewater treatment, and public water supplies. Solid waste treatment/ disposal has best potential role in reducing GHG emissions. In general, therefore, it seems that reduction strategies for the main contributor should be prior to other categories and lead to best practice for managing GHG emissions, especially considering annual budgets.

This study characterized PM and VOC emissions from cow dung combustion in a controlled experiment. Dung from grass-fed cows was dried and combusted using a dual cone calorimeter. Heat fluxes of 10, 25, and 50 kW/m² were applied. The concentrations of PM and VOCs were determined using a dust spectrometer and gas chromatography/mass spectrometry, respectively. PM and VOC emission factors were much higher for the lower heat flux, implying a fire ignition stage. When the heat flux was 50 kW/m², the CO₂ emission factor was highest and the PM and VOC emission factors were lowest. Particle concentrations were highest in the 0.23-0.3-μm size range at heat fluxes of 25 kW/m² and 50 kW/m². Various toxic VOCs including acetone, methyl ethyl ketone, benzene, and toluene were detected at high concentrations. Although PM and VOC emission factors at 50 kW/m² were lower, they were high enough to cause extremely high indoor air pollution. The characteristics of PM and VOC emissions from cow dung combustion indicated potential health effects of indoor air pollution in developing countries.

Due to rapid industrialization and population growth uncontrolled release of heavy metals are entered into the waters. Among these heavy metals Pb(II) is one of the major toxic metal and in recent years the production and consumption of lead is increasing worldwide. Pb(II) can be entered to aqueous streams from several industries and can enter into the humans food chain through drinking water and crop irrigation. Lead can causes severe damage to the kidney, nervous system, reproductive system, liver and brain. The permissible level for lead in drinking water is 0.05 mg/l. Thus in recent years a number of methods and materials were developed to removal Pb(II) from aqueous solutions. Among these material bio-chars obtained from plant materials have gained special attention due to their low-cost and abundant nature. In present investigation we have developed magnetic bio-char composite from pine bark. Pine trees are wide spread throughout the South Korea and the bark from pine tree has no commercial use and is available as waste. Thus we have utilized this waste inexpensive material from preparing bio-char composite. The pin bark obtained was initially made into fine powder and washed several times with water and was filtered. To this powder an appropriate amounts of nitrate salts of cobalt and iron dissolved in ethanol solution was added and stirred for 15 minutes. This solution was oven dried at 70℃ and this was further calcined at 900℃ in nitrogen atmosphere. As obtained material was washed several times with water and dried in oven over night. This was used as adsorbent for treating lead contaminated aqueous solutions. As obtained bio-char composite was used to remove Pb(II) from aqueous solutions. Various parameters influencing Pb(II) removal like initial pH, contact time and initial concentration were studied. Effect of pH on Pb(II) removal was studied in the pH range from 2-8 at Pb(II) concentration 10 mg/L using an adsorbent dose of 300 mg. At below pH 3 a lower percent removal was observed whereas above pH 4>90% removal was observed. Further effect of contact time on Pb(II) removal was studied from time range between 10-180 min. Two kinetic models pseudo-first, pseudo-second-order models were used to evaluated the kinetic data and found that the data was better fitted to the pseudo-second-order model. From the overall results it was found that as prepared magnetic bio-char composite prepared from pin bark waste was effective and economic for treating Pb(II) contaminated aqueous solutions.

In order to raise awareness about environmental protection, people are paying more attention now-a-days in reusing wastepaper. As a result, most countries in the world have already made significant progress related to wastepaper recycling technology. The aim of this study was to investigate the effects of slow-release nitrogen fertilizer (SRNF) on the growth of radish plants. Wastepaper was deinked by alkaline solution and SRNF was produced from fertilizer impregnated wastepaper, which applied to an experimental plot compared with a urea fertilized plot. The plant height and total chlorophyll content of the radishes were higher while they were treated with SRNF than with urea. Some agronomic and chemical components were also observed and significant differences between the two fertilizers were found. When the soil was treated with SRNF, the pH, organic matter and total nitrogen content were higher than in the soil which was treated with urea.

In this study, we attempted to solubilize protein in slaughter blood (SB) using ultrasonic technology. The application of ultrasonic technology can make enzymatic degradation of SB more effective, which has no comparable alternative for treatment. The SB was homogenized by grinding it for 10 minutes at 10,000 rpm as a pretreatment for preventing its clotting, and then ultrasonic treatment was attempted to solubilize protein in SB. To maximize the efficiency of ultrasonic treatment for SB, the optimum condition of ultrasonic frequency (UF) was determined to be 20 kHz. To optimize the operation conditions of ultrasonification with 20 kHz of frequency, we used response surface methodology (RSM) based on ultrasonic density (UD) and ultrasonification time (UT). The solubilization rate (SR) of protein (%) was calculated to be 101.304 - 19.4205 X1 + 0.0398X2 + 7.9411X12 + 0.0001X22 + 0.0455X1X2. From the results of the RSM study, the optimum conditions of UD and UT were determined at 0.5 W/mL and 22 minutes, respectively, and SB treated under these conditions was estimated to have a 95% SR. Also, experimentally, a 95.53% SR was observed under same conditions, accurately reflecting the theoretical prediction of 95%.

Bisphenol A (BPA) is an estrogenic endocrine disrupter. However, depending on a way of treatment, the harmful effects of BPA have not been confirmed. Also, trans-generational effects of BPA on male reproduction are still controversial. Because the reabsorption of testicular fluid in the efferent ductules (ED) and initial segment (IS) is important for sperm maturation, the present study was designed to determine trans-generational effect of BPA administrated orally on expression of water transport-related molecules in the mouse ED and IS. Ethanol-dissolved BPA was diluted in water to be 100 ng (low), 10 ㎍ (medium), and 1 ㎎/㎖ water (high). BPA-containing water was provided for two generations. Expression of ion transporters and water channels in the ED and IS were measured by relative real-time PCR analysis. In the ED, BPA treatment caused expressional increases of carbonic anhydrase II, cystic fibrosis ransmembrane regulator, Na+/K+ ATPase α1 subunit, and aquaporin (AQP) 1. No change of Na+/H+ exchange (NHE) 3 expression was detected. BPA treatment at medium dose resulted in an increase of AQP9 expression. In the IS, the highest expressional levels of all molecules tested were observed in medium-dose BPA treatment. Generally, high-dose BPA treatment resulted in a decrease or no change of gene expression. Fluctuation of NHE3 gene expression by BPA treatment at different concentrations was detected. These findings suggest that trans-generational exposure to BPA, even at low dose, could affect gene expression of water-transport related molecules. However, such effects of BPA would be differentially occurred in the ED and IS.

To effects of sex maturation in olive flounder by regulating long photoperiod, gonadal development and GTH mRNA expression in the pituitary were investigated. Photoperiod was treated natural photoperiod and long photoperiod (15L:9D) conditions from September 2011 to March 2012. The results showed that natural photoperiodic group showed a higher gonadosomatic index (GSI) than long photoperiodic group during the spawning season (March 2012). The histological analysis of ovarian tissue showed that natural photoperiod group of ovaries contained vitellogenic oocytes, but long photoperiod group of ovaries mainly contained perinucleolus staged oocyte and oil-drop staged oocytes. The FSH mRNA of olive flounder, under natural photoperiod group, showed a significantly higher expression but no significant difference under long photoperiod group. The LHβ mRNA showed a significantly higher expression only under natural photoperiod group. These results may suggest that long photoperiodic information regulates secretion of pituitary FSH and LH and maintain early growing stage of gonadal development in this species.

Olive flounder (Paralichthys olivaceus) is one of the commercial important flatfish species in Korea. The ocular signal transduction pathway is important in newly hatched flounders because it is closely involved in the initial feeding phase thus essential for survival during the juvenile period. However, the study of gene expression during ocular development is incomplete in olive flounder. Therefore we examined the expression analysis of specifically induced genes during the development of the visual system in newly hatched flounders. We searched ocular development-involved gene in the database of expressed sequence tags (ESTs) from olive flounder eye and this gene similar to arrestin with a partial sequence homology. Microscopic observation of retinal formation corresponded with the time of expression of the arrestin gene in the developmental stage. These results suggest that arrestin plays a vital role in the visual signal transduction pathway of the retina during ocular development. The expression of arrestin was strong in the ocular system during the entirety of the development stages. Our findings regarding arrestin have important implications with respect to its biological role and evolution of G-protein coupled receptor (GPCR) signaling in olive flounder. Further studies are required on the GPCR-mediated signaling pathway and to decipher the functional role of arrestin.

아스타잔틴을 포함하는 수중유형 형태의 나노에멀젼을 고압균질기를 이용하여 제조하였다. 이에 유화조건, 유화제 종류, 유화제 농도 그리고 아스타잔틴 농도에 따라 최적화를 되었다. 나노에멀젼의 안정성은 제타포텐셜, FF-SEM, 입도분석기, 색차계를 이용하여 측정하였다. 제조된 아스타잔틴 나노에멀젼의 입도는 160 ~190 nm로 균일하였으며 레시친에 의한 나노에멀젼 보다 glyceryl citrate/lactate/linoleate/oleate에 의한 나노에멀젼이 더욱 안정하고 균일한 입도분포를 가졌다. 아스타잔틴의 봉입도는 HPLC, FF-SEM을 이용하여 확인 하였으며, 제형 구성 후 보관조건에서의 안정도 및 제타 포텐셜 값도 -41의 우수한 결과를 나타내었다.

Cathepsins are members of the multigene family of lysosomal cysteine proteinases and have regulated function in several life processes. The potential role of cathepsin F cysteine gene was expected as protease in the yolk processing mechanism during early developmental stage, but expression analysis was unknown after fertilization. The alignment analysis showed that amino acid sequence of cathepsin F from olive flounder liver expressed sequence tag (EST) homologous to cathepsin F of other known cathepsin F sequences with 87－98% identity. In this study, we examined the gene expression analysis of cathepsin F in various tissues at variety age flounder. Tissue distribution of the cathepsin F mRNA has been shown to be ubiquitous and constitutive pattern regardless of age in each group, although derived from cDNA library using liver sample. The mRNA level of cathepsin F more increased as developmental proceed during embryogenesis and early developmental stage, especially increased in the blastula, hatching stage and 3 days post hatching (dph). As a result, it may suggest that the proteolysis of yolk proteins (YPs) has been implicated as a mechanism for nutrient supply during early larval stages in olive flounder.

Pluripotent cells are categorized as either "naive" or "primed" based upon their pluripotent status. According to previous studies, embryonic stem cells and embryonic germ cells are identified as naive pluripotent stem cells and epiblast stem cells are identified as primed pluripotent stem cells. In a permissive species such as the mouse, naive and primed pluripotent stem cells can be derived from embryos without genetic manipulations. In non-permissive species such as humans and pigs, primed pluripotent cells are only established from embryos. However, previous studies have shown that the embryonic germ cells of non-permissive species share similar morphology and features with naive pluripotent cells. For these reasons porcine embryonic germ cells (pEGCs) may provide a useful cell source for comparative studies on naive pluripotent cells in non-permissive species. In this study, we attempted to establish and characterize porcine embryonic germ cells. Consequently, an embryonic germ cell line was derived from the genital ridges of a porcine dpc 30 fetus in media containing bFGF. This cell line displayed a dome-shaped colony morphology. The cell line was maintained in a stable condition over an extended time period and was able to differentiate into the three germ layers in vitro. Pluripotency markers such as OCT4, SOX2, NANOG and SSEA4 were expressed in these pEGCs. Similar with pESCs, Mek/Erk signaling pathway were activated by bFGF in the cultured pEGCs. In conclusion, we were able to successfully derive embryonic germ cells from genital ridges of a porcine fetus. Unlikely naive pluripotent cells such as mESCs, pluripotency of pEGCs were regulated by Mek/Erk signaling pathway activated by bFGF. This cell line could potentially be used as naive pluripotent cell source for comparative study with porcine embryonic stem cells and other pluripotent cell lines. As porcine pluripotent cells, pEGCs could be useful candidates for preliminary studies of human disease as well as a source for generating transgenic animals.

Cleft palates with or without cleft lip is one of the most common congenital craniofacial defects in dogs. It has been reported that monogenic autosomal recessive inheritance caused this defect in this species. However, here, we aimed to report cleft palate in a cloned dog. A fibroblast cell line was established from skin tissues of an eight-year-old German shepherd dog. Blood was collected from oocyte donor dogs, and serum progesterone concentration was measured by chemiluminescence enzyme immunoassay method. Ovulation was determined when serum progesterone results reached 5-10 ng/ml, and in vivo matured oocytes were collected surgically about 72 hr after ovulation. Donor cells were cultured with Dulbecco’s modified Eagle medium supplemented with 10% (v/v) fetal bovine serum until confluence. An in vivo matured oocyte was enucleated, and a donor cell was injected into the perivitelline space. The oocyte-cell couplet was electrically fused, and chemically activated. Reconstructed embryos were transferred to an oviduct of a recipient. Pregnancy diagnosis was performed 27 days after the embryo transfer, and ultrasonography of fetal heart beat, and rectal temperature and serum progesterone value of recipient was monitored until the day of delivery. Microsatellite analysis was performed using genomic DNA of cell donor, clones, and oocyte donors. As results, a total of 74 cloned embryos were transferred to five recipients, and one recipient diagnosed as pregnant with two fetuses by ultrasonography and radiology. Caesarean section was performed on day 58 after embryo transfer due to a decreased heart beat of a fetus, which was lower than 180. Two cloned puppies with 640g and 320g of birth weight were delivered safety, but the small one was born with a cleft palate. Microsatellite analysis results of both clones were identical with the cell donor. Cleft palate of the clone was surgically corrected on day 40 after birth. To our knowledge, there has been no report about cleft palate in cloned dogs, and also, no report about clones with different phenotype of cleft palate in dogs. Therefore, this study can give a clue of cleft palate in dogs, which might not be a genetic cause. Further studies about aberrant epigenetic reprogramming in those clones are needed.

Olive flounder (Paralichthys olivaceus) is a most important aquaculture species in Korea. Like most marine fishes, olive flounders are stomachless at first feeding and aquired gastric function during the metamorphosis, so food was mainly digested by pancreatic enzyme from first feeding to premetamorphosis. However, comprehensive analysis of pancreatic and gastric digestive enzyme of olive flounder at early developmental period is still unclear. In the expression study of pancreatic and gastric digestive enzyme by real-time PCR at early developmental stage, pancreatic enzyme such as chymotrypsinogen 2, preproelastase 2 and 4, pancreatic protein somatomedin-B domain (PPSB) mRNA expression were initiated at first feeding and strongly expressed in the pancreas developmental stage, while gastric digestive enzyme signal was not at all detected during same period. Although, trypsinogens were secreted from pancreas and have similar amino acid sequence, trypsinogen 3 expression induction was detected both pancreas and stomach developmental stage, while trypsinogen 2 expression was significantly increased only post-metamorphosis period. Pepsinogen mRNA expression was only detected at metamorphosis according to stomach differentiation. Lipid digestive enzyme, lipase and intestine fatty acid binding protein 1 (I-FABP 1), were already reached a certain level at beginning of hatching and more increased during early developmental stage and then gradually decreased before metamorphosis. These results suggested that feed ingestion of olive flounder was exclusive charged by pancreatic digestive enyme, lipid digestive enzyme and trypsinogen 3 from first feeding and then fully swiched by gastric digestive enzyme and trypsinogen 2 from metamorphosis period.

Soluble-NSF attachment protein receptor (SNARE) proteins play a role in vesicle fusion, exocytosis, and intracellular trafficking in neuronal cells as well as in fertilization and embryogenesis. We investigated the expression patterns of two SNARE proteins, SNAP-25 and synaptotagmin VII (SytVII), and their regulation by pregnant mare serum gonadotropin (PMSG) during mouse ovarian follicular development. Ovaries were obtained at 0, 12, 24, 36, and 48 h post-PMSG injection of immature mice. SNAP-25 and SytVII mRNA expression levels increased gradually in a time-dependant manner. However, protein levels revealed different patterns of expression, suggesting different translational regulation following PMSG stimulation. SNAP-25 and SytVII expression was closely associated with thickening of the granulosa cell (GC) layer and follicle morphological changes from a flattened to a cuboidal shape. To explore follicle stimulating hormone receptor (FSHR)-mediated regulation of their expression, GCs from preantral follicles were cultured to examine the effects of FSHR siRNA knockdown. FSHR siRNA abolished upregulation of the SNAREs in both PMSG and FSH-stimulated GCs. This abolished gene expression was rescued by adding dibutyryl cyclic AMP to the cultures. These results suggest that SNAP-25 and SytVII expression is regulated via the FSHR-cAMP pathway during follicular development.

Use of nature-derived matrices of a part of body tissues has been used to repair damaged tissues in practical terms. Recently, the same idea has also been applied to regenerate whole organs including the heart, liver, lung, and pancreas etc. Thus, so-called bio-artificial organ technology becomes a promising way of overcoming the lack of donor organs and immune rejections in organ transplantation if we can obtain recipient stem cells. Although the regenerated heart in vitro so far may demonstrate some typical organ's responses in vitro and vivo, it is still far from a fully functional organ for transplantation. We initiated a study to look at changes occurring during the generation of bio-artificial organ using the mouse model. Adult hearts were dissected out and perfused for acellularization with SDS-containing buffer and washed several times. Enzymatic treatment also evaluated the acellular purity by isolating genomic DNA and total RNA before and after DNase and RNase treatments. For recellularization, differentiating H9C2 cell or cells derived from P19 EC cells along with mesenchymal stem cells were seeded on the finally obtained heart matrix several times before submerging culture for generating the heart. Histological analyses revealed that complete removal of cellular components. The intensive staining of alcian blue (pH 1.5 and 2.5) suggests that acid mucopolysaccharides, glycocomponents and sulfate-containing saccharides are widely spread within the heart matrix. There was little DNA and RNA in the heart matrix after the enzymatic treatments as judging by the DAPI or PI staining. Cell seeding and subsequent submerging culture showed substantial heart tissue development as evidenced by immunocytochemistry and RT-PCR in the recellularized and grown heart. From these results, we suggest that each procedure for bio-artificial organ has to be carefully examined to improve the entire process.

One of the most effective and safe therapeutic methods for treating vitiligo, mixed autologous keratinocytes (KCs) and melanocytes (MCs) cultures have been used for autologous cell transplantation. However, the present transplantation method is faced with a problem that may require a large amount of skin tissue and keratinocytes have limited culture potency. We have found previously that human adipose derived stromal cells (hASCs) from aspirated fat tissue could be used in place of KCs and sufficient amounts of hASCs for transplantation could be obtained by small amount of aspirated fat tissue. The present investigation was determined the effect of ASCs on ex vivo expansion MCs for transplantation. In addition, we examined for a preservation conditions of MCs which have reported low recovery rates and a slowdown in growth after cryopreservation. Various conditions including ASCs ratio, incubation period, and additive materials for MCs cultivation was determined to improve the expansion ability of MCs. The growth rate of MCs colony was elevated 6.85 folds compared the previous conditions. These MCs showed a specific expression of immature melanocyte protein, Trp-2, but did not express the mature melanocyte proteins and markers (c-kit, CD133, and etc.) of mesenchymal stem cells that represents in ASCs feeder. Results in cryopreservation experiments were determined a preservation medium for MCs showing an increased recovery rates after thawing. The characteristics of MCs after cryopreservation using a designed medium were indicated consistent morphology and immunophenotype. In conclusion, ASCs as a feeder could be used in place of keratinocytes for ex vivo expansion of MCs. For clinical trial for vitiligo patients, efficiency experiments in preclinical state should be followed.

Pluripotent stem cells are cells that have a self-renewal capacity and the ability to differentiate into all lineages. These cells can be divided into naive- and primed-state pluripotent stem cells according to their pluripotent state. Only the naive state comprises a full pluripotency or ground state that contributes to germ-line transmission. Naive states are found in specific permissive strains or species, such as 129, C57BL/6 and BALB/C in mice. However, a number of attempts have been made to derive naive-state pluripotent stem cell lines from non-permissive species, including humans and pigs, using various exogenous factors including GSK3β and MEK inhibitors (2i), LIF, hypoxic conditions and up-regulation of Oct4 or Klf4. Therefore, in this study we investigated whether a naive pluripotent stem cell line could be derived from porcine embryonic fibroblasts (PEFs) via previously reported factors. Our mouse embryonic stem cell (mESC)-like cell lines expressed the pluripotency markers Oct4, Sox2 and Nanog and a stable mESC-like morphology for more than 50 passages. In addition, these cell lines could be sequentially reprogrammed into mESC-like induced pluripotent stem (iPS) cells from secondary or tertiary fibroblast-like cells differentiated from mESC-like iPS cells by addition of doxycycline (DOX), LIF and 2i. Our results suggest that, as a non-permissive species, porcine stem cells can be induced into mESC-like iPS cells from PEFs by various exogenous factors, including continuous transgene expression, 2i and LIF. However, further work that aims to effectively induce the activation of endogenous transcription factors is necessary to derive authentic naive-state pluripotent porcine stem cells.

Anthocyanins are the largest group of water-soluble pigments in the plant kingdom. They are widely distributed in the human diet through crops, beans, fruits, vegetables and red wine. The specific health effects that anthocyanins might have in vivo are not known, although there are several possibilities related to obesity, cardiovascular disease, and cancer. In this study we used human subcutaneous adipose mesenchymal stem cells (hADSC) and mouse subcutaneous adipose mesenchymal stem cells (mADSC) to evaluate the effects of anthocyanins. And we examined the effect of cell activity and adipocyte differentiation by Cyanidin-3-O-glucoside (C3G), Delphinidin-3-ß-D-glucoside (D3G) that are among the anthocyanin family and black soybean extract. Using MTT assay method, we tested cellular metabolic activity. In mADSC, cell activity is significantly decreased by C3G and D3G (50 uM, 100 uM, and 200 uM), and black soybean extract (100 ug/ml and 200 ug/ml). In hADSC, cell activity is significantly decreased only by C3G (50 uM, 100 uM and 200 uM) unlike in mADSC. Cell activity is significantly increased of 100 uM D3G and black soybean extract (50 ug/ml, 100 ug/ml and 200 ug/ml). In mADSC, 50 uM C3G promoted differentiation into adipocyte but no effect in other concentration. D3G suppressed the differentiation of mADSC at 100 uM and 200 uM. 50 ug/ml black soybean extract promoted differentiation of mADSC, but 200 ug/ml black soybean extract suppressed differentiation. In hADSC, 50 uM, 100 uM and 200 uM C3G suppressed differentiation. 100 uM D3G promoted differentiation into adipocyte, but 200uM D3G suppressed it. Black soybean extract suppressed the differentiation into adipocyte at 50 ug/ml, 100 ug/ml and 200 ug/ml. These data showed that the responsibility to the C3G and D3G were different between hADSC and mADSC. Interestingly the responsibility to the black soybean extract was similar between hADSC and mADSC. Based on them, it is suggested that there are species-specificity to the cellular responsibility to the anthocyanins in subcutaneous ADSC.