• Title/Summary/Keyword: Single Cell

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In Vitro Aggregation and Culture of Mouse Embryos (생쥐 배의 시험관내 응집과 배양)

  • 이상진;정길생
    • Korean Journal of Animal Reproduction
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    • v.8 no.1
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    • pp.29-35
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    • 1984
  • These experiments were carried out to obtain basic information necessary for in vitro culture of aggregated mouse embryos. Inbred ICR mice were used to obtain embryos. The zona pellucida was removed by placing the embryos in Whittingham's medium containing 0.5% protease for about 5-10minutes at 37$^{\circ}C$. Total 263 pairs of 2-, 4- and 8-cell zona free mouse embryos were subjected to aggregation by physical pressure and cultured in Whittingham's medium under the gas phase of 5% CO2 in air at 37$^{\circ}C$ for 24 to 60 hours. The results obtained in these experiments were summarized as follows: 1. Time needed for fusion of 2-, 4- and 8-cell embryos were 0-3, 0-3 and 0-3 hours, respectively and average time needed for in vitro development of 2-, 4- and 8-cell embryos after aggregation to morula and blastocyst were 42, 30 and 13.5 hours, and 51, 39 and 27 hours, respectively. 2. Of total 263 pairs of naked embryos, 227 were firmly aggregated together and the rats of aggregation in 2-, 4- and 8-cell embryos were 71.8, 88.3 and 97.0%, respectively. 3. The rates of aggregated pairs which obtained from 2-, 4- and 8-cell embryos developed to morula were 96.7, 95.6 and 96.9%, respectively, and embryos developed to blastocysts were 88.5, 89.7 and 90.8%, respectively. 4. Conspicuous differences in size of volume and inner cell masses between single and double blastocysts were observed. Although a single blastocolic cavity was formed in most double blastocysts, several formed two distinct cavities from the very beginning.

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Fabrication and Characterization of Composite LSCF-Ag Cathode for Solid Oxide Fuel Cells using Electron Beam Irradiation Process

  • Kang, Hyun Suk;Jung, Yung-Min;Song, Rak-Hyun;Peck, Dong-Hyun;Park, ChangMoon;Lee, Byung Cheol
    • Bulletin of the Korean Chemical Society
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    • v.35 no.10
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    • pp.2969-2973
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    • 2014
  • A new process to fabricate a composite LSCF-Ag cathode material for SOFCs by electron beam (e-beam) irradiation process has been suggested for operation under intermediate temperature range of $600-700^{\circ}C$. A composite LSCF-Ag cathode with uniformly coated Ag nanoparticles on the surface of the LSCF material was prepared by a facile e-beam irradiation method at room temperature. The morphology of the composite LSCF-Ag material was analyzed using a TEM, FE-SEM, and EDS. The prepared composite LSCF-Ag material can play a significant role in increasing the electro-catalytic activities and reducing the operating temperature of SOFCs. The performance of a tubular single cell prepared using the composite LSCF-Ag cathode, YSZ electrolyte and a Ni/YSZ anode was evaluated at reduced operating temperature of $600-700^{\circ}C$. The micro-structure and chemical composition of the single cell were investigated using a FE-SEM and EDS.

Single Cell Transcriptomic Re-analysis of Immune Cells in Bronchoalveolar Lavage Fluids Reveals the Correlation of B Cell Characteristics and Disease Severity of Patients with SARS-CoV-2 Infection

  • Chae Won Kim;Ji Eun Oh;Heung Kyu Lee
    • IMMUNE NETWORK
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    • v.21 no.1
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    • pp.10.1-10.13
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    • 2021
  • The coronavirus disease 2019 (COVID-19) pandemic (severe acute respiratory syndrome coronavirus 2) is a global infectious disease with rapid spread. Some patients have severe symptoms and clinical signs caused by an excessive inflammatory response, which increases the risk of mortality. In this study, we reanalyzed scRNA-seq data of cells from bronchoalveolar lavage fluids of patients with COVID-19 with mild and severe symptoms, focusing on Ab-producing cells. In patients with severe disease, B cells seemed to be more activated and expressed more immunoglobulin genes compared with cells from patients with mild disease, and macrophages expressed higher levels of the TNF superfamily member B-cell activating factor but not of APRIL (a proliferation-inducing ligand). In addition, macrophages from patients with severe disease had increased pro-inflammatory features and pathways associated with Fc receptor-mediated signaling, compared with patients with mild disease. CCR2-positive plasma cells accumulated in patients with severe disease, probably because of increased CCL2 expression on macrophages from patients with severe disease. Together, these results support the hypothesis that different characteristics of B cells might be associated with the severity of COVID-19 infection.

QoS Constrained Optimization of Cell Association and Resource Allocation for Load Balancing in Downlink Heterogeneous Cellular Networks

  • Su, Gongchao;Chen, Bin;Lin, Xiaohui;Wang, Hui;Li, Lemin
    • KSII Transactions on Internet and Information Systems (TIIS)
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    • v.9 no.5
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    • pp.1569-1586
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    • 2015
  • This paper considers the optimal cell association and resource allocation for load balancing in a heterogeneous cellular network subject to user's quality-of-service (QoS) constraints. We adopt the proportional fairness (PF) utility maximization formulation which also accommodates the QoS constraints in terms of minimum rate requirements. With equal resource allocation this joint optimization problem is either infeasible or requires relaxation that yields a solution which is difficult to implement. Nevertheless, we show that this joint optimization problem can be effectively solved without any priori assumption on resource allocation and yields a cell association scheme which enforces single BS association for each user. We re-formulated the joint optimization problem as a network-wide resource allocation problem with cardinality constraints. A reweighted heuristic l1-norm regularization method is used to obtain a sparse solution to the re-formulated problem. The cell association scheme is then derived from the sparsity pattern of the solution, which guarantees a single BS association for each user. Compared with the previously proposed method based on equal resource allocation, the proposed framework results in a feasible cell association scheme and yields a robust solution on resource allocation that satisfies the QoS constraints. Our simulations illustrate the impact of user's minimum rate requirements on cell association and demonstrate that the proposed approach achieves load balancing and enforces single BS association for users.

A Study on Radio Resource Management for Multi-cell SC-FDMA Systems (다중셀 SC-FDMA를 위한 무선자원 관리기법에 관한연구)

  • Chung, Yong-Joo
    • Journal of Korea Society of Industrial Information Systems
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    • v.15 no.4
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    • pp.7-15
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    • 2010
  • This study proposes a rad o resource management scheme to maximize the performance of the LTE(Long Term Evolution) uplink, using SC-FDMA(Single Carrier-Frequency Division Multiple Access). Rather than the single-cell SC-FDMA system the existing studies are mainly concerning, this study focuses on multi-cell system which needs considering the interaction among cells. Radio resource management is divided into two phases, planning and operation phases. The former is for the master eNB(e-NodeB) to allocate RBs(radio bearer) to eNB, the latter for eNB to assign RBs to the mobiles in the cell. For each phase, an optimization model and greedy algorithm are proposed. Optimization models aim to maximize the system performance while satisfying the constraints for both QoS and RB continuity. The greedy algorithms, like generic ones, move from a solution to a neighboring one having the best objective value among neighboring ones. From the numerous numerical experiments, the performance and characteristics of the algorithms are analyzed. This study is expected to play a volunteering role in radio resource management for the multi-cell SC-FDMA system.

Identification of Cell Type-Specific Effects of DNMT3A Mutations on Relapse in Acute Myeloid Leukemia

  • Seo-Gyeong Bae;Hyeoung-Joon Kim;Mi Yeon Kim;Dennis Dong Hwan Kim;So-I Shin;Jae-Sook Ahn;Jihwan Park
    • Molecules and Cells
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    • v.46 no.10
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    • pp.611-626
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    • 2023
  • Acute myeloid leukemia (AML) is a heterogeneous disease caused by distinctive mutations in individual patients; therefore, each patient may display different cell-type compositions. Although most patients with AML achieve complete remission (CR) through intensive chemotherapy, the likelihood of relapse remains high. Several studies have attempted to characterize the genetic and cellular heterogeneity of AML; however, our understanding of the cellular heterogeneity of AML remains limited. In this study, we performed single-cell RNA sequencing (scRNAseq) of bone marrow-derived mononuclear cells obtained from same patients at different AML stages (diagnosis, CR, and relapse). We found that hematopoietic stem cells (HSCs) at diagnosis were abnormal compared to normal HSCs. By improving the detection of the DNMT3A R882 mutation with targeted scRNAseq, we identified that DNMT3A-mutant cells that mainly remained were granulocyte-monocyte progenitors (GMPs) or lymphoid-primed multipotential progenitors (LMPPs) from CR to relapse and that DNMT3A-mutant cells have gene signatures related to AML and leukemic cells. Copy number variation analysis at the single-cell level indicated that the cell type that possesses DNMT3A mutations is an important factor in AML relapse and that GMP and LMPP cells can affect relapse in patients with AML. This study advances our understanding of the role of DNMT3A in AML relapse and our approach can be applied to predict treatment outcomes.

Single-Cell RNA Sequencing of Bone Marrow Mesenchymal Stem Cells from the Elderly People

  • Dezhou Zhu;Jie Gao;Chengxuan Tang;Zheng Xu;Tiansheng Sun
    • International Journal of Stem Cells
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    • v.15 no.2
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    • pp.173-182
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    • 2022
  • Background and Objectives: Bone marrow mesenchymal stem cells (BMSCs) show considerable promise in regenerative medicine. Many studies demonstrated that BMSCs cultured in vitro were highly heterogeneous and composed of diverse cell subpopulations, which may be the basis of their multiple biological characteristics. However, the exact cell subpopulations that make up BMSCs are still unknown. Methods and Results: In this study, we used single-cell RNA sequencing (scRNA-Seq) to divide 6,514 BMSCs into three clusters. The number and corresponding proportion of cells in clusters 1 to 3 were 3,766 (57.81%), 1,720 (26.40%), and 1,028 (15.78%). The gene expression profile and function of the cells in the same cluster were similar. The vast majority of cells expressed the markers defining BMSCs by flow cytometry and gene expression analysis. Each cluster had at least 20 differentially expressed genes (DEGs). We conducted Gene Ontology enrichment analysis on the top 20 DEGs of each cluster and found that the three clusters had different functions, which were related to self-renewal, multilineage differentiation and cytokine secretion, respectively. In addition, the function of the top 20 DEGs of each cluster was checked by the National Center for Biotechnology Information gene database to further verify our hypothesis. Conclusions: This study indicated that scRNA-Seq can be used to divide BMSCs into different subpopulations, demonstrating the heterogeneity of BMSCs.

The development of murine recombinant single-chain variable domain fragment (ScFv) specific to acute non-lymphocytic leukemia (ANLL) cell line HL60 (인간의 급성 비임파성 백혈암세포(HL60)의 표면항원에 결합하는 재조합 single-chain Fv (ScFv)의 개발)

  • Kim, Cheol Hong;Han, Seung Hee;Kim, Hyeong Min;Han, Jae Yong;Lim, Myeong Woon;Kim, Jin-Kyoo
    • Korean Journal of Microbiology
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    • v.51 no.2
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    • pp.115-125
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    • 2015
  • A monoclonal antibody AP64 IgM binds to human acute nonlymphocytic leukemia (ANLL) cell line HL60 and also cross-reacts with the homologous antigen in a rat ANLL cell. This antibody mediated by complement, has leukemia a suppression effect. In this study, we generated a recombinant single-chain variable domain fragment (ScFv) which were derived from $V_H$ and $V_L$ cDNA of AP64 IgM-secreting hybridoma by RT-PCR. The two variable regions were joined with a single 15 amino acid linker $(G_4S)_3$. This recombinant ScFv was expressed as a single polypeptide chain from Escherichia coli BMH 71-18. The recombinant ScFv was purified by applying the periplasmic extract to $Ni^+$-NTA-agarose affinity column and detected with westernblot. The purified recombinant ScFv recognized a surface antigen (about 30 kDa) of HL60 cell line which is the same antigen detected by parental AP64 IgM. But the affinity of ScFv for a surface antigen of HL60 was lower than that of the parental AP64 IgM, which needs to be further improved. Overall, the recombinant ScFv specific to HL60 might be a useful bioreagent for either diagnostic or therapeutic purposes.

Comparison between Single and Co-culture of Adipocyte and Muscle Cell Lines in Cell Morphology and Cytosolic Substances (지방과 근육 세포주의 단독 및 공동배양을 통한 세포형태학 및 세포물질 비교 연구)

  • Choi, Chang-Weon;Cho, Won-Mo;Yeon, Seong-Heum;HwangBo, Soon;Song, Man-Kang;Park, Sung-Kwon;Baek, Kyung-Hoon
    • Journal of Animal Science and Technology
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    • v.54 no.2
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    • pp.103-109
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    • 2012
  • Present study was performed to investigate the effect of single and co-culture of adipocyte and muscle cell lines on cell differentiation. 3T3-L1 (adipocyte) and L6 (muscle) cell lines were single-cultured on the condition of 10% fetal bovine serum (FBS)/Dulbeco's modified eagle's medium (DMEM) for 48 h followed by culture within 5% FBS/DMEM as a growth media. Then, the growth media was replaced by differentiation media composed of 2% FBS/DMEM without additives in single- or co-culture of the 3T3-L1 and the L6 cells to induce differentiation of both cell types. In co-culture system, the 3T3-L1 and the L6 cells were grown in separated places by being seeded on a $0.4{\mu}m$ insert membrane and on the bottom of 6 well plate, respectively. Cell differentiation was measured using morphological investigation and cytosolic analysis of glycerol-3-phosphate dehydrogenase (GPDH; for 3T3-L1) and creatine kinase (CK; for L6). Based on the GPDH results, the presence of L6 cells did not stimulate 3T3-L1 differentiation showing more differentiation of 3T3-L1 cells in the single-culture compared to the co-culture condition. In contrast, 3T3-L1 cells in the co-culture promoted differentiation of L6 cells. Enzymatic analysis supported this result showing that 3T3-L1 cells showed statistically (P<0.05) higher GPDH activity in the single-culture than the co-culture, whereas CK results of L6 cells were vice versa (P<0.05). Overall, present results may indicate that co-culture system is more reliable and precise technique compared to single-culture. Further studies on several co-culture trials including different media conditions, supplementation of differentiating substances, molecular biological analysis, etc. should be required to obtain practical and fundamental mass data.

Preparation of Porous Carbon Fiber by Using MgO Powder and Its Characteristics of Catalysts for Fuel Cell (MgO를 이용한 다공성 탄소 섬유 제조 및 이를 이용한 연료전지용 촉매 특성)

  • Nam, Kidon;Kim, Sang-Kyung;Lim, Seongyop;Peck, Donghyun;Lee, Byoungrok;Jung, Doohwan
    • Korean Chemical Engineering Research
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    • v.46 no.6
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    • pp.1142-1147
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    • 2008
  • Nano-structured porous carbon fiber(PCF) for the catalyst supports of the direct methanol fuel cell (DMFC) were prepared from the mesophase pitch by using the nano-MgO powders. Specific surface area of the PCFs was $8{\sim}58m^2/g$ and surface pore structures had almost meso pore diameter of 10~20 nm which were depending on the amount of MgO spheres. Aqueous reduction method was used to load 60 wt% PtRu on the prepared PCF supports. The electro-oxidation activity and single cell performance of the 60 wt% Pt-Ru catalysts were measured by cyclic voltammetry and unit cell test. The performances of these catalysts increased by 5~10% compared with one of commercial catalyst.