• Title/Summary/Keyword: surface markers

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New Isolation Technique and Culture System for Clinical Applications of Human Amniotic Epithelial Stem Cells (인간태반양막유래 상피줄기세포의 임상적용을 위한 새로운 세포분리 및 배양 기술)

  • Woo, Sang-Kyu;Jo, Jung-Yoon;Shin, Il-Seob;Kang, Sung-Keun;Ra, Jeong-Chan
    • Development and Reproduction
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    • v.13 no.4
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    • pp.271-280
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    • 2009
  • Human placenta is abundant source of adult stem cells. Especially, amniotic epithelial cells have stem cell characteristics, expressing surface markers normally present on embryonic stem cells and germ cells. However, culturing and expanding amniotic epithelial cells in vitro without feeder cells are difficult due to endogenous characteristics of epithelial cells. In the present study, amniotic epithelial cells are isolated and proliferated in several passages by applying dithiothreitol and a Rho-associated kinase inhibitor in culture media. The cultured amniotic epithelial cells showed the epithelial and stem cell characteristics. In conclusion, human placenta-derived amniotic epithelial stem cells can be a major source of stem cells for medical treatment of various diseases without any controversial issues.

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Genetic characteristics of the Korean isolate KI -1 of Toxoplasma gondii

  • LIN Aifen;SHIN Eun-Hee;KIM Tae-Yun;PARK Jae-Hwan;GUK Sang-Mee;CHAI Jong-Yil
    • Parasites, Hosts and Diseases
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    • v.43 no.1 s.133
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    • pp.27-32
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    • 2005
  • Toxoplasma gondii tachyzoites were isolated from an ocular patient in the Republic of Korea and maintained in the laboratory (designated KI-1). In the present study, its genotype was determined by analyzing dense granule antigen 6 (GRA6) gene and surface antigen 2 (SAG2) gene as typing markers. Digestion of the amplification products of GRA6 and of the 5' and 3' ends of SAG2, respectively, with Mse I, Sau3A I, and Hha I, revealed that KI-1 is included in the genotype I, which includes the worldwide virulent RH strain. In addition, when the whole sequences of the coding regions of SAG1, rhoptry antigen 1 (ROP1), and GRA8 genes of KI-1 were compared with those of RH, minor nucleotide polymorphisms and amino acid substitutions were identified. These results show that KI-1 is a new geographical strain of T. gondii that can be included in the genotype I.

Bacterial strains isolated from Jeotgal (salted seafood) induce maturation and cytokine production in mouse bone marrow-derived dendritic cells (마우스 골수 유래 수지상세포의 성숙과 사이토카인 생산에 대한 젓갈 분리균의 효과 연구)

  • Moon, Sun-Young;Park, Eun-Jin;Joo, Hong-Gu
    • Korean Journal of Veterinary Research
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    • v.54 no.3
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    • pp.139-146
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    • 2014
  • Jeotgal (salted seafood) has been one of major fermented foods in Korea for long time. Although there are many studies about Jeotgal in various aspects of food, its immunological importance on hosts has not been elucidated yet. In this study, we investigated if several bacteria isolated from Jeotgal may modulate the function of dendritic cells (DCs), powerful antigen-presenting cells equipped with special immunological capabilities. 4 Jeotgal bacteria were selected as representatives and used for experiments. To treat viable DCs, those bacteria were killed at $60^{\circ}C$ for 30 min. The viability of DCs treated with Jeotgal bacteria was verified and two isolates significantly induced high production of interleukin-12, a representative cell-mediated cytokine of DCs. Surface activation and maturation markers (MHC class II, CD40, CD86) of DCs were analyzed by flow cytometer. In addition, the treated DCs showed significantly high lymphocyte stimulatory capability compared to control DCs based on allogeneic mixed lymphocyte reactions. These observations suggest that Jeotgal isolates can function as immunostimulating bacteria in hosts, like Lactobacillus. Taken together, these experimental evidences may broaden the use of Jeotgal isolates in immunological fields in addition to as a fermented food.

Chondrogenesis of Mesenchymal Stem Cell Derived form Canine Adipose Tissue

  • Lee, Byung-Joo;Wang, Soo-Geun;Seo, Cheol-Ju;Lee, Jin-Chun;Jung, Jin-Sup;Lee, Ryang-Hwa
    • Proceedings of the KSLP Conference
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    • 2003.11a
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    • pp.183-183
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    • 2003
  • Background and Objectives : Cartilage reconstruction is one of medical issue in otolaryngology. Tissue engineering is presently being utilized in part of cartilage repair. Sources of cells for tissue engineering are chondrocyte from mature cartilage and bone marrow mesenchymal stem cells that are able to differentiate into chondrocyte. Recent studies have shown that adipose tissue have mesenchymal stem cells which can differentiate into adipogenic, chondrogenic myogenic osteogenic cells and neural cell in vitro. In this study, we have examined chondrogenic potential of the canine adipose tissue-derived mesenchymal stem cell(ATSC). Materials and Methods : We harvested canine adipose tissue from inguinal area. ATSCs were enzymatically released from canine adipose tissue. Under appropriate culture conditions, ATSCs were induced to differentiate into the chondrocyte lineages using micromass culture technique. We used immunostain to type II collagen and toluidine blue stain to confirm chondrogenic differentiation of ATSCs. Results : We could isolate ATSCs from canine adipose tissue. ATSCs expressed CD29 and CD44 which are specific surface markers of mesenchymal stem cell. ATSCs differentiated into micromass that has positive response to immunostain of type II collagen and toluidine blue stain. Conclusion : In vitro, ATSCs differentiated into cells that have characteristic cartilage matrix molecules in the presence of lineage-specific induction factors. Adipose tissue may represent an alternative source to bone marrow-derived MSCs.

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Morphological evaluation during in vitro chondrogenesis of dental pulp stromal cells (영구치 치수 기질세포를 이용한 연골 분화 및 분화 시기에 따른 형태학적 변화)

  • Chung, Choo-Ryung;Kim, Ha-Na;Park, Yeul;Kim, Min-Jeong;Oh, Young-Ju;Shin, Su-Jung;Choi, Yoon-Jeong;Kim, Kyung-Ho
    • Restorative Dentistry and Endodontics
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    • v.37 no.1
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    • pp.34-40
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    • 2012
  • Objectives: The aim was to confirm the stem cell-like properties of the dental pulp stromal cells and to evaluate the morphologic changes during in vitro chondrogenesis. Materials and Methods: Stromal cells were outgrown from the dental pulp tissue of the premolars. Surface markers were investigated and cell proliferation rate was compared to other mesenchymal stem cells. Multipotency of the pulp cells was confirmed by inducing osteogenesis, adipogenesis and chondrogenesis. The morphologic changes in the chondrogenic pellet during the 21 day of induction were evaluated under light microscope and transmission electron microscope. TUNEL assay was used to evaluate apoptosis within the chondrogenic pellets. Results: Pulp cells were CD90, 105 positive and CD31, 34 negative. They showed similar proliferation rate to other stem cells. Pulp cells differentiated to osteogenic, adipogenic and chondrogenic tissues. During chondrogenesis, 3-dimensional pellet was created with multi-layers, hypertrophic chondrocyte-like cells and cartilage-like extracellular matrix. However, cell morphology became irregular and apoptotic cells were increased after 7 day of chondrogenic induction. Conclusions: Pulp cells indicated mesenchymal stem cell-like characteristics. During the in vitro chondrogenesis, cellular activity was superior during the earlier phase (within 7 day) of differentiation.

Inhibitory Effect of Artemisiae Asiaticae Herba on Degranulation, Production of Cytokine, and $Fc{\varepsilon}RI$ Expression in RBL-2H3 Cells (애엽(艾葉)의 RBL-2H3 비만세포 탈과립, cytokine 생산 및 $Fc{\varepsilon}RI$ 유전자 발현에 미치는 영향)

  • Ban, Hak Jin;Park, Dong II;Kang, Kyung Hwa
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.26 no.6
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    • pp.915-921
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    • 2012
  • Artemisiae Asiaticae Herba (AAH) has been used to remedy of symptoms such as bleeding, dysmenorrhea, eczema and itchy skin in Oriental Medicine. In this study, we investigated the protective effect of AAH on allergic response. The effect of AAH was analyzed by ELISA and RT-PCR in RBL-2H3 cells. We investigated cell viability, ${\beta}$-hexosaminidase and histamine as markers of degranulation, production of IL-4 and TNF-${\alpha}$, and gene expression of HDC2, cytokines and $Fc{\varepsilon}RI$ ${\alpha}{\beta}{\gamma}$ subunit. We found that AAH suppressed ${\beta}$-hexosaminidase and histamine release, the production of IL-4 and TNF-${\alpha}$ in RBL-2H3 by the anti-DNP IgE plus DNP-HSA stimulation. AAH also significantly decreased cytokine mRNA expressions, such as IL-$1{\beta}$, IL-2, IL-3, IL-4, IL-5, IL-6, IL-12, IL-13, TNF-${\alpha}$, and GM-CSF, and increased cytokine mRNA expressions of IL-10 in RBL-2H3. In addition, AAH suppressed mRNA expression of $Fc{\varepsilon}RI$ ${\alpha}{\beta}{\gamma}$ subunit on cell surface. Our results indicate that AAH protects against allergic response and exerts an anti-inflammatory effect through the inhibition of degranulation and production of cytokines and expression of $Fc{\varepsilon}RI$ ${\alpha}{\beta}{\gamma}$ subunit.

The effects of cryopreservation on human dental pulp-derived mesenchymal stem cells

  • Tomlin, Allison;Sanders, Michael B;Kingsley, Karl
    • Biomaterials and Biomechanics in Bioengineering
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    • v.3 no.2
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    • pp.105-114
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    • 2016
  • The purpose of this study is to evaluate the effects of cryopreservation on dental pulp-derived stem cells (DPSC) viability over a period of three years. Dental pulp-derived stem cells were isolated and cultured from thirty-one healthy teeth. DPSC isolates were assessed for doubling-time and baseline viability prior to cryopreservation and were assessed again at three time points; one week (T1), 18 months (T2), and 36 months (T3). DPSC can be grouped based on their observed doubling times; slow (sDT), intermediate (iDT), and rapid (rDT). Viability results demonstrated all three types of DPSC isolates (sDT, iDT and rDT) exhibit time-dependent reductions in viability following cryopreservation, with the greatest reduction observed among sDT-DPSCs and the smallest observed among the rDT-DPSC isolates. Cryopreserved DPSCs demonstrate time-dependent reductions in cellular viability. Although reductions in viability were smallest at the initial time point (T1) and greatest at the final time point (T3), these changes were markedly different among DPSC isolates with similar doubling times (DTs). Furthermore, the analysis of various DPSC biomarkers - including both intracellular and cell surface markers, revealed differential mRNA expression. More specifically, the relative high expression of Sox-2 was only found only among the rDT isolates, which was associated with the smallest reduction in viability over time. The expression of Oct4 and NANOG were also higher among rDT isolates, however, expression was comparatively lower among the sDT isolates that had the highest reduction in cellular viability over the course of this study. These data may suggest that some biomarkers, including Sox-2, Oct4 and NANOG may have some potential for use as biomarkers that may be associated with either higher or lower cellular viability over long-term storage applications although more research will be needed to confirm these findings.

Quickly Map Renewal through IPM-based Image Matching with High-Definition Map (IPM 기반 정밀도로지도 매칭을 통한 지도 신속 갱신 방법)

  • Kim, Duk-Jung;Lee, Won-Jong;Kim, Gi-Chang;Choi, Yun-Soo
    • Korean Journal of Remote Sensing
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    • v.37 no.5_1
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    • pp.1163-1175
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    • 2021
  • In autonomous driving, road markings are an essential element for object tracking, path planning and they are able to provide important information for localization. This paper presents an approach to update and measure road surface markers with HD maps as well as matching using inverse perspective mapping. The IPM removes perspective effects from the vehicle's front camera image and remaps them to the 2D domain to create a bird-view region to fit with HD map regions. In addition, letters and arrows such as stop lines, crosswalks, dotted lines, and straight lines are recognized and compared to objects on the HD map to determine whether they are updated. The localization of a newly installed object can be obtained by referring to the measurement value of the surrounding object on the HD map. Therefore, we are able to obtain high accuracy update results with very low computational costs and low-cost cameras and GNSS/INS sensors alone.

Effects of a Powder Formulation of Streptomyces cameroonensis on Growth and Resistance of Two Cocoa Hybrids from Cameroon against Phytophthora megakarya (Causal Agent of Black Pod Disease)

  • Aristide, Dzelamonyuy;Martial, Tene Tayo Paul;Ruth, Ngotcho Ngassam Esther;Grace, Lele Brenda;Ebenezer, Foka Tatiekam;Flore, Magni Pacha Tatiana;Thaddee, Boudjeko
    • Journal of Microbiology and Biotechnology
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    • v.32 no.2
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    • pp.160-169
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    • 2022
  • In the present study we evaluated the efficacy of a bioformulation of Streptomyces cameroonensis for control of black pod disease in cocoa and enhancement of seedling growth. The formulation developed using talc powder and cassava starch as carriers showed high shelf-life of 1.07 × 106 CFU/g after six months storage at 4℃. The formulation was tested for inhibition of spore germination in Phytophthora megakarya and showed 100% inhibition at 10% (w/v) of formulation. To determine the efficacy of the formulation, we performed an in planta assay in the greenhouse on two hybrids of cocoa seedlings, the tolerant SNK413 × (♂) T79/467 and the susceptible UPA 134× (♂) SCA 12. Detached leaf assay showed a significant reduction in the disease severity index of about 67% for the tolerant hybrid and 55% for the susceptible hybrid compared to non-treated plants. A significant enhancement in stem length, leaf surface area and root weight was observed. Analysis of biochemical markers of defense showed a significant increase in total polyphenol, flavonoid, and total protein contents. There was also significant upregulation of PR-proteins such as chitinases, peroxidases and β-1, 3-glucanases following treatment of both tolerant and susceptible hybrids, though with a higher level of synthesis in the tolerant hybrids. A significant increase was also observed in polyphenol oxidase activities in plants treated with the formulation. This work demonstrated the stability and effectiveness of the S. cameroonensis powder formulation in suppressing black pod disease in cocoa and subsequently enhancing the growth of seedlings.

Monoclonal antibody K312-based depletion of pluripotent cells from differentiated stem cell progeny prevents teratoma formation

  • Park, Jongjin;Lee, Dong Gwang;Lee, Na Geum;Kwon, Min-Gi;Son, Yeon Sung;Son, Mi-Young;Bae, Kwang-Hee;Lee, Jangwook;Park, Jong-Gil;Lee, Nam-Kyung;Min, Jeong-Ki
    • BMB Reports
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    • v.55 no.3
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    • pp.142-147
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    • 2022
  • Human pluripotent stem cells (PSCs) have been utilized as a promising source in regenerative medicine. However, the risk of teratoma formation that comes with residual undifferentiated PSCs in differentiated cell populations is most concerning in the clinical use of PSC derivatives. Here, we report that a monoclonal antibody (mAb) targeting PSCs could distinguish undifferentiated PSCs, with potential teratoma-forming activity, from differentiated PSC progeny. A panel of hybridomas generated from mouse immunization with H9 human embryonic stem cells (hESCs) was screened for ESC-specific binding using flow cytometry. A novel mAb, K312, was selected considering its high stem cell-binding activity, and this mAb could bind to several human induced pluripotent stem cells and PSC lines. Cell-binding activity of K312 was markedly decreased as hESCs were differentiated into embryoid bodies or by retinoic acid treatment. In addition, a cell population negatively isolated from undifferentiated or differentiated H9 hESCs via K312 targeting showed a significantly reduced expression of pluripotency markers, including Oct4 and Nanog. Furthermore, K312-based depletion of pluripotent cells from differentiated PSC progeny completely prevented teratoma formation. Therefore, our findings suggest that K312 is utilizable in improving stem cell transplantation safety by specifically distinguishing residual undifferentiated PSCs.