• 제목/요약/키워드: Stem Cell Medium

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Platycarya strobilacea S. et Z. Extract Has a High Antioxidant Capacity and Exhibits Hair Growth-promoting Effects in Male C57BL/6 Mice

  • Kim, Eun Jin;Choi, Joo Yeon;Park, Byung Cheol;Lee, Bog-Hieu
    • Preventive Nutrition and Food Science
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    • v.19 no.3
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    • pp.136-144
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    • 2014
  • This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. This study was conducted to evaluate the effects of Platycarya strobilacea S. et Z. (PSE) extract on mouse hair growth and to determine the mechanism of action of PSE. PSE was purchased and its antioxidant activities, such as electron donating ability, total polyphenol content, and flavonoid content were tested. Toxicity during topical treatment was determined by the CCK-8 assay, a cell viability test. Fifteen 4-week-old male C57BL/6 mice were assigned to receive one of three treatments: dimethyl sulfoxide (negative control), minoxidil (positive control) or PSE. Test materials were topically applied to the shaved dorsal skin of each mouse daily for 3 weeks. After 21 days, we observed skin tissue hair follicle morphology and length, mast cell number, and stem cell factor (SCF) expression using hematoxylin and eosin (H&E), toluidine blue, and immunohistochemical staining, respectively. Furthermore, the expression of cytokines involved in hair growth [i.e., insulin-like growth factor (IGF)-1, keratinocyte growth factor (KGF), and transforming growth factor (TGF)-${\beta}1$] was determined by PCR. PSE was found to have very high antioxidant activity. The cell viability rate of PSE-treated mice was markedly higher than that of mice in the control group. We also observed an increase in hair follicle length, strong SCF staining, and a decrease in mast cell number in the PSE group. In addition, PSE-treated mice had higher IGF-1 and KGF expression and lower TGF-${\beta}1$ expression than mice in the minoxidil-treated group. These results suggest that topical application of PSE promotes hair growth by intensifying SCF, suppressing mast cell production, and increasing hair growth-promoting cytokine expression.

Analysis of Stromal Cells Developed from Cord Blood CD34+ Cells (제대혈 CD34+ 세포에서 유래된 지지세포의 분석)

  • Ryu, Kyung-Ha;Park, Se-Jin;Kim, Kyung Hyo;Seoh, Ju-Young;Khan, Mohammad;Shin, Hee-Young;Ahn, Hyo-Seop
    • IMMUNE NETWORK
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    • v.1 no.1
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    • pp.87-94
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    • 2001
  • Background: Cytokine-mediated ex vivo expansion has been proposed as a means of increasing the number of cord blood (CB) hematopoietic stem cells for transplantation. As well as stem cell number, stromal cells are necessary for functional maturation of hematopoiesis. The purpose of this study was to analyze the development of stromal cells during ex vivo expansion of CB $CD34^+$ cells. Methods : $CD34^+$ cells were purified from CB by magnetic bead selection. The levels of of interleukin-3, interleukin-$1{\beta}$, interleukin-6, granulocyte macrophagecolony stimulating factor and tumor necrosis factor-${\alpha}$ were measured in culture supernatants on 0, 1, 2, and 3 weeks, using ELISA techniques. CB $CD34^+$ cells were expanded in Iscoves modified Dulbeccos medium in the presence of several cytokines. The expression of E-selectin, vascular cell adhesion molecule-1, intercellular adhesion molecule-1, platelet/endothelial cell adhesion molecule-1, von Willebrand factor, vimentin, and CD14 in newly developed stromal cells was examined by immunocytochemical method. Relevant extracellular matrix (ECM) proteins and proper cytokines were also assayed for the most suitable condition for expansion of stromal cells. Results: Several cytokines were found to have been produced by CB $CD34^+$ cells as well as bone marrow-derived $CD34^+$ cells. During ex vivo expansion of CB $CD34^+$ cells, stromal cells appeared in the culture by day 4 and expanded over the following 7-10 days before being confluent by day 2 1. These cells expressed surface markers characteristic of cells of endothelial lineage. Furthermore, these stroaml cells also expanded effectively when treated with thrombopoietin+flt-3 ligand+stem cell factor+leukemia inhibitory factor or 0.1% poly-L-lysine-coated wells. Conclusion: Stromal cells were developed during ex vivo expansion of CB $CD34^+$ cells and that this development could be enhanced further by treating the stromal cells with cytokines or ECM.

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Use of Human Adipose Tissue as a Source of Endothelial Cells (혈관내피세포 채취의 원천으로 인간 지방조직의 활용)

  • Park, Bong-Wook;Hah, Young-Sool;Kim, Jin-Hyun;Cho, Hee-Young;Jung, Myeong-Hee;Kim, Deok-Ryong;Kim, Uk-Kyu;Kim, Jong-Ryoul;Jang, Jung-Hui;Byun, June-Ho
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.32 no.4
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    • pp.299-305
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    • 2010
  • Purpose: Adipose tissue is located beneath the skin, around internal organs, and in the bone marrow in humans. Its main role is to store energy in the form of fat, although it also cushions and insulates the body. Adipose tissue also has the ability to dynamically expand and shrink throughout the life of an adult. Recently, it has been shown that adipose tissue contains a population of adult multipotent mesenchymal stem cells and endothelial progenitor cells that, in cell culture conditions, have extensive proliferative capacity and are able to differentiate into several lineages, including, osteogenic, chondrogenic, endothelial cells, and myogenic lineages. Materials and Methods: This study focused on endothelial cell culture from the adipose tissue. Adipose tissues were harvested from buccal fat pad during bilateral sagittal split ramus osteotomy for surgical correction of mandibular prognathism. The tissues were treated with 0.075% type I collagenase. The samples were neutralized with DMEM/and centrifuged for 10 min at 2,400 rpm. The pellet was treated with 3 volume of RBC lysis buffer and filtered through a 100 ${\mu}m$ nylon cell strainer. The filtered cells were centrifuged for 10 min at 2,400 rpm. The cells were further cultured in the endothelial cell culture medium (EGM-2, Cambrex, Walkersville, Md., USA) supplemented with 10% fetal bovine serum, human EGF, human VEGF, human insulin-like growth factor-1, human FGF-$\beta$, heparin, ascorbic acid and hydrocortisone at a density of $1{\times}10^5$ cells/well in a 24-well plate. Low positivity of endothelial cell markers, such as CD31 and CD146, was observed during early passage of cells. Results: Increase of CD146 positivity was observed in passage 5 to 7 adipose tissue-derived cells. However, CD44, representative mesenchymal stem cell marker, was also strongly expressed. CD146 sorted adipose tissue-derived cells was cultured using immuno-magnetic beads. Magnetic labeling with 100 ${\mu}l$ microbeads per 108 cells was performed for 30 minutes at $4^{\circ}C$ a using CD146 direct cell isolation kit. Magnetic separation was carried out and a separator under a biological hood. Aliquous of CD146+ sorted cells were evaluated for purity by flow cytometry. Sorted cells were 96.04% positivity for CD146. And then tube formation was examined. These CD146 sorted adipose tissue-derived cells formed tube-like structures on Matrigel. Conclusion: These results suggest that adipose tissue-derived cells are endothelial cells. With the fabrication of the vascularized scaffold construct, novel approaches could be developed to enhance the engineered scaffold by the addition of adipose tissue-derived endothelial cells and periosteal-derived osteoblastic cells to promote bone growth.

Comparative Analysis about the Effect of Isolated Phosphatidylcholine and Sodium Deoxycholate for the Viability of Adipocyte (Phosphatidylcholine과 Sodium Deoxycholate가 지방세포 생존에 미치는 영향의 비교 분석)

  • Rha, Eun-Young;Kang, Jo-A;Lee, Jung-Ho;Oh, Deuk-Young;Seo, Je-Won;Moon, Suk-Ho;Ahn, Sang-Tae;Rhie, Jong-Won
    • Archives of Plastic Surgery
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    • v.37 no.5
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    • pp.531-534
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    • 2010
  • Purpose: Lipobean$^{(R)}$s, widely used in lipodissolving techniques, contain phosphatidylcholine and sodium deoxycholate as its main substances. They have been approved only as medication for liver disease by the FDA. However, they have been used under various clinical settings without exact knowledge of its action mechanism. The authors designed an in vitro study to analyze the effects of different concentrations of phosphatidylcholine and sodium deoxycholate on adipocytes and other types of cells. Methods: Human adipose-derived stem cell were cultured and induced to differentiate into adipocytes. Fibroblasts extracted from human inferior turbinate tissue, and MC3T3-E1 osteoblast lines were cultured. Phosphatidylcholine solution dissolved with ethanol was applied to the culture medium at differing concentrations (1, 4, 7, 10 mg/mL). The sodium deoxycholate solution dissolved in DMSO applied to the medium at differing concentrations (0.07, 0.1. 0.4. 0.7 mg/mL). Cells were dispersed at a concentration of $5{\times}10^3$ cells/well in 24 well plates, and surviving cells were calculated 1 day after the application using a CCK-8 kit. Results: The number of surviving cells of adipocytes, fibroblasts and osteoblasts decreased as the concentration of sodium deoxycholate increased. However, all types of cells that had been processed in a phosphatidylcholine showed a cell survival rate of over 70% at all concentrations. Conclusion: This study shows that sodium deoxycholate is the more major factor in destroying adipocytes, and it is also toxic to the other cells. Therefore, we conclude that care must be taken when using Lipobean$^{(R)}$s as a method of reducing adipose tissue, for its toxicity may destroy other nontarget cells existing in the subcutaneous tissue layer.

In vitro regeneration and the change of anatomical appearance in Poncirus trifoliata RAFIN. (탱자(Poncirus trifoliata RAFIN.)의 기내 재분화 및 조직학적 특성)

  • 박민희;이현화;장현규;이숙영;김홍섭
    • Korean Journal of Plant Resources
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    • v.12 no.2
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    • pp.107-119
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    • 1999
  • In this study, the induction regeneration of callus from immature embryo in trifoliata orange (Poncirus trifoliata RAFIN.) were accomplished. The embryogenic calli were induced from the immature embryo derived from seed when the calli were irradiated for 16hr at about 2,000 Lux in $\frac{1}{2}$ MS medium supplemented with 3% sucrose, and 44.4$\mu$M BA. Regeneration to whole plants was the most successful in MS medium containing 5.0$\mu$M BA. The yellowish callus was developed at 2 to 3 weeks of culture and the callus was changed from yellow to green at 5 to 6 weeks culture. In vitro regeneration was directly induced from embryogenic callus in MS medium containing 3% sucrose and 5.0$\mu$M BA. Multishoot was formed at 16 weeks culture. Moreover, when the root-formed plantlet was transplanted to soil, they grew to a whole plant. The compact cultured-cells were observed by light microscope after 4 weeks of cultivation and the embryogenic clumps were formed about the 5 weeks. At the same time, the neighboring cells were liquefied. In addition, differentiation of leaf and stem from the callus was observed after 12 weeks. The developed oil sacs and the profacicular cambium of the immature leaf were observed after 18 weeks. Therefore, we can see the considerable changes of cell arrangements according to the developmental stages of calli from trifoliata orange.

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Evaluation of Combined Contrast Agent using N-(p-maleimidophenyl) Isocyanate Linker-mediated Synthesis for Simultaneous PET-MRI (동시 PET-MRI를 위한 N-(p-maleimidophenyl) isocyanate linker-매개 합성을 이용한 복합 조영제의 평가)

  • Lee, Gil-Jae;Lee, Hwun-Jae;Lee, Tae-Soo
    • Journal of the Korean Society of Radiology
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    • v.16 no.2
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    • pp.103-113
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    • 2022
  • In this paper, a combined 18F-FDG(fluorodeoxyglucose) and MNP(magnetic nanoparticles) contrast agent was synthesized using N-(p-maleimidophenyl) isocyanate as the crosslinker for use in simultaneous PET-MRI scans. PET-MRI images were acquired and evaluated before and after injection of the combined contrast imaging agent (18F-FDG labeled MNP) from a glioma stem cell mouse model. After setting the region of interest (ROI) on each acquired image, the area of the lesion was calculated by segmentation. As a result, the PET image was larger than the MRI. In particular, the simultaneous PET-MRI images showed accurate lesions along with the surrounding soft tissue. The mean and standard deviation values were higher in the MRI images alone than in the PET images or the simultaneous PET-MRI images, regardless of whether the contrast agent was injected. In addition, the simultaneous PET-MRI image values were higher than for the PET images. For PSNR experiments, the original image was PET Image using 18F-FDG, MRI using MNPs, and MRI without contrast medium, and the target image was simultaneous PET-MRI image using 18F-FDG labeled MNPs contrast medium. As a result, all of them appeared significantly, suggesting that the 18F-FDG labeled MNPs contrast medium is useful. Future research is needed to develop an agent that can simultaneously diagnose and treat through SPECT-MRI imaging research that can use various nuclides.

Characteristics of bioethanol production using sweet sorghum juice as a medium of the seed culture (단수수 착즙액이용 배양종균의 바이오에탄올 생산 특성 연구)

  • Cha, Young-Lok;Moon, Youn-Ho;Yu, Gyeong-Dan;Lee, Ji-Eun;Choi, In-Seung;Song, Yeon-Sang;Lee, Kyeong-Bo
    • Journal of the Korean Applied Science and Technology
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    • v.33 no.4
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    • pp.627-633
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    • 2016
  • Sweet sorghum [Sorghum bicolor (L)] is one of the major crops for biofuels such as sugarcane and sugar beet which raw materials rich in saccharide. Sweet sorghum juice was extracted from the stem. It's composed of fermentable sugars such as glucose, fructose and sucrose. Ethanol from the extracted sweet sorghum juice can be easily produced by yeast fermentation process. Sweet sorghum juice is consisted of not only sugars but also various nutrients like nitrogen and phosphate. For commercial production of bioethanol, seed culture is one of the important parts of fermentation, so that optimal culture medium should be selected for the reduction of processing costs. In this study, sweet sorghum juice was estimated as a culture medium for seed culture of cellulosic bioethanol. For the comparison of cultures with various substrates, it used YPD including each 5 g/L yeast extract and peptone, sweet sorghum juice and hydrolyzed Miscanthus was taken part in the culture with 2%, 5% and 10% sugar conditions. Based on media of YPD and sweet sorghum juice, cell-mass concentration was obtained maximum more than $2.5{\times}10^8CFU/mL$ after 24 h of cultivation. Consequently sweet sorghum juice is suitable for the cell culture with more than $1.0{\times}10^8CFU/mL$ after 12 h of cultivation. This can be used as a culture medium for the cellulosic bioethanol industry.

Isolation and Identification of Antifungal Compounds from $Bacillus$ $subtilis$ C9 Inhibiting the Growth of Plant Pathogenic Fungi

  • Islam, Md. Rezuanul;Jeong, Yong-Tae;Lee, Yong-Se;Song, Chi-Hyun
    • Mycobiology
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    • v.40 no.1
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    • pp.59-65
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    • 2012
  • Antagonistic microorganisms against $Rhizoctonia$ $solani$ were isolated and their antifungal activities were investigated. Two hundred sixteen bacterial isolates were isolated from various soil samples and 19 isolates were found to antagonize the selected plant pathogenic fungi with varying degrees. Among them, isolate C9 was selected as an antagonistic microorganism with potential for use in further studies. Treatment with the selected isolate C9 resulted in significantly reduced incidence of stem-segment colonization by $R.$ $solani$ AG2-2(IV) in Zoysia grass and enhanced growth of grass. Through its biochemical, physiological, and 16S rDNA characteristics, the selected bacterium was identified as $Bacillus$ $subtilis$ subsp. $subtilis$. Mannitol (1%) and soytone (1%) were found to be the best carbon and nitrogen sources, respectively, for use in antibiotic production. An antibiotic compound, designated as DG4, was separated and purified from ethyl acetate extract of the culture broth of isolate C9. On the basis of spectral data, including proton nuclear magneric resonance ($^1H$ NMR), carbon nuclear magneric resonance ($^{13}C$ NMR), and mass analyses, its chemical structure was established as a stereoisomer of acetylbutanediol. Application of the ethyl acetate extract of isolate C9 to several plant pathogens resulted in dose-dependent inhibition. Treatment with the purified compound (an isomer of acetylbuanediol) resulted in significantly inhibited growth of tested pathogens. The cell free culture supernatant of isolate C9 showed a chitinase effect on chitin medium. Results from the present study demonstrated the significant potential of the purified compound from isolate C9 for use as a biocontrol agent as well as a plant growth promoter with the ability to trigger induced systemic resistance of plants.

Altered Gene Expression Profiles in the Lungs of Streptozotocin-induced Diabetic Mice

  • Kim, Jung-Hyun;Rasaei, Roya;Park, Sujin;Kim, Ji-Young;Na, Sunghun;Hong, Seok-Ho
    • Development and Reproduction
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    • v.24 no.3
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    • pp.197-205
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    • 2020
  • Diabetes mellitus is a common heterogeneous metabolic disorder, characterized by deposition of extracellular matrix, oxidative stress, and vascular dysfunction, thereby leading to gradual loss of function in multiple organs. However, little attention has been paid to gene expression changes in the lung under hyperglycemic conditions. In this study, we found that diabetes inuced histological changes in the lung of streptozotocin-induced diabetic mice. Global gene expression profiling revealed a set of genes that are up- and down-regulated in the lung of diabetic mice. Among these, expression of Amigo2, Adrb2, and Zbtb16 were confirmed at the transcript level to correlate significantly with hyperglycemia in the lung. We further evaluated the effect of human umbilical cord-derived perivascular stem cells (PVCs) on these gene expression in the lung of diabetic mice. Our results show that administration of PVC-conditioned medium significantly suppressed Amig2, Adrb2, and Zbtb16 upregulation in these mice, suggesting that these genes may be useful indicators of lung injury during hyperglycemia. Furthermore, PVCs offer a promising alternative cell therapy for treating diabetic complications via regulation of gene expression.

The relationship between reactive oxygen species, DNA fragmentation, and sperm parameters in human sperm using simplified sucrose vitrification with or without triple antioxidant supplementation

  • Juanpanich, Theesit;Suttirojpattana, Tayita;Parnpai, Rangsun;Vutyavanich, Teraporn
    • Clinical and Experimental Reproductive Medicine
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    • v.49 no.2
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    • pp.117-126
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    • 2022
  • Objective: This study examined whether the addition of triple antioxidants (3A)-10 µM acetyl-L-carnitine, 10 µM N-acetyl-L-cysteine, and 5 µM α-lipoic acid-in freezing-thawing medium during human sperm cryopreservation using the sucrose vitrification (SuV) and liquid nitrogen vapor (Vapor) techniques could improve post-thaw survival of spermatozoa. Methods: We analyzed 30 samples from healthy human sperm donors. Each sample was allocated into one of five groups: fresh control, SuV, SuV+3A, Vapor, and Vapor+3A. The sperm motility, morphology, viability, intracellular and extracellular reactive oxygen species (ROS) levels, and sperm DNA fragmentation (SDF) were evaluated. Results: The cryopreserved spermatozoa had significantly reduced percentages of motility (p<0.05) and viability (p<0.05). Antioxidant supplementation non-significantly improved these parameters (p>0.05). No significant differences were found in sperm morphology between the fresh and frozen-thawed groups (p>0.05). After freezing, the extracellular ROS levels in the frozen-thawed groups were significantly higher (p<0.05) than in the fresh group. However, we did not find any differences in intracellular ROS parameters among these groups (p>0.05). The SDF was higher in the SuV and Vapor groups than in the fresh group, but without statistical significance (p=0.075 and p=0.077, respectively). Conclusion: Cryopreservation had detrimental effects on sperm motility, viability, and extracellular ROS levels, without changing the morphology or intracellular ROS levels. Antioxidant supplementation was slightly effective in preventing SDF in frozen-thawed spermatozoa.