• Title/Summary/Keyword: Single-Cell Analysis

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Dissection of Cellular Communication between Human Primary Osteoblasts and Bone Marrow Mesenchymal Stem Cells in Osteoarthritis at Single-Cell Resolution

  • Ying Liu;Yan Chen;Xiao-Hua Li;Chong Cao;Hui-Xi Zhang;Cui Zhou;Yu Chen;Yun Gong;Jun-Xiao Yang;Liang Cheng;Xiang-Ding Chen;Hui Shen;Hong-Mei Xiao;Li-Jun Tan;Hong-Wen Deng
    • International Journal of Stem Cells
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    • v.16 no.3
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    • pp.342-355
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    • 2023
  • Background and Objectives: Osteoblasts are derived from bone marrow mesenchymal stem cells (BMMSCs) and play important role in bone remodeling. While our previous studies have investigated the cell subtypes and heterogeneity in osteoblasts and BMMSCs separately, cell-to-cell communications between osteoblasts and BMMSCs in vivo in humans have not been characterized. The aim of this study was to investigate the cellular communication between human primary osteoblasts and bone marrow mesenchymal stem cells. Methods and Results: To investigate the cell-to-cell communications between osteoblasts and BMMSCs and identify new cell subtypes, we performed a systematic integration analysis with our single-cell RNA sequencing (scRNA-seq) transcriptomes data from BMMSCs and osteoblasts. We successfully identified a novel preosteoblasts subtype which highly expressed ATF3, CCL2, CXCL2 and IRF1. Biological functional annotations of the transcriptomes suggested that the novel preosteoblasts subtype may inhibit osteoblasts differentiation, maintain cells to a less differentiated status and recruit osteoclasts. Ligand-receptor interaction analysis showed strong interaction between mature osteoblasts and BMMSCs. Meanwhile, we found FZD1 was highly expressed in BMMSCs of osteogenic differentiation direction. WIF1 and SFRP4, which were highly expressed in mature osteoblasts were reported to inhibit osteogenic differentiation. We speculated that WIF1 and sFRP4 expressed in mature osteoblasts inhibited the binding of FZD1 to Wnt ligand in BMMSCs, thereby further inhibiting osteogenic differentiation of BMMSCs. Conclusions: Our study provided a more systematic and comprehensive understanding of the heterogeneity of osteogenic cells. At the single cell level, this study provided insights into the cell-to-cell communications between BMMSCs and osteoblasts and mature osteoblasts may mediate negative feedback regulation of osteogenesis process.

Influence of co-culturing muscle satellite cells with preadipocytes on the differentiation of adipocytes and muscle cells isolated from Korean native cattle

  • Choi, Chang Weon
    • Korean Journal of Agricultural Science
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    • v.45 no.4
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    • pp.715-723
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    • 2018
  • The present study was done to investigate the effect of co-culturing muscle satellite cells (MSCs) and intramuscular preadipocytes (IPs) on the differentiation of adipocytes and muscle cells isolated from Korean native cattle. MSCs and IPs were single-cultured in 10% fetal bovine serum/Dulbecco's modified Eagles medium (FBS/DMEM) for 48 h followed by culturing in 5% FBS/DMEM as the growth media. Then, the growth media was replaced by differentiation media composed of 2% FBS/DMEM without any additives for the single- or co-culture of muscle cells and intramuscular adipocytes to induce the differentiation of both cell types. Cell differentiation was measured by morphological investigation and cytosolic enzyme analysis of glycerol-3-phosphate dehydrogenase (GPDH) for the adipocytes and creatine kinase (CK) for the muscle cells. In the morphological test, the presence of muscle cells did not stimulate adipocyte differentiation showing more differentiation of the adipocytes in the single-culture compared to the co-culture condition. However, the differentiation of muscle cells was promoted by adipocytes in the co-culture. The results of the enzymatic analysis were highly associated with the morphological results with a statistically higher GPDH activity (p < 0.05) appearing in the single-culture than in the co-culture, whereas the opposite was true for the CK activity of the muscle cells (p < 0.05). By manipulating in vivo the milieu using a co-culture, we could detect the difference in the rate of cell differentiation and suggest that a co-culture system is a more reliable and precise technique compared to a single-culture. Further studies on various co-culture trials including supplementation of differentiating substances, gene expression analysis, etc. should be done to obtain practical and fundamental data.

ANALYSIS OF CHROMOSOME ABERRATION, SISTER CHROMATID EXCHANGE, MICRONUCLEI AND SINGLE CELL GEL ELECTROPHORESIS IN HUMAN LYMPHOCYTES EXPOSED IN VITRO TO BISPHENOL A AND DIETHYLSTILBESTROL

  • Kim, Byeong-Mo;Chung, Hai-Won
    • Proceedings of the Korean Society of Toxicology Conference
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    • 2001.10a
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    • pp.176-176
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    • 2001
  • Endocrine disruptors have been implicated in carcinogenesis in animal studies, but carcinogenetic effects on human remain controversial. In order to examine the genotoxicity of two common endocrine disruptors, Bisphenol A and Diethylstilbestrol, cytogenetic endpoints including chromosome aberration(CA), sister chromatid exchange (SCE), micronuclei (MN) analyses and DNA damage by single cell gel electrophoresis (SCGE) were assayed.(omitted)

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신경경로의 정보처리에 대한 전기적 특성 연구

  • 박상희;이명호
    • 전기의세계
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    • v.28 no.8
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    • pp.66-71
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    • 1979
  • This paper describes electrical analysis of the information processing of the nervous system. A general-purpose electrical neuronal model for simulating the electrical activity in a single nerve cell and in small groups of nerve cell has constructed. This model consists of two basic electronic modules to represent respectively a "cell body" and an "axon (with synapses)", together with various related appurtenances. The primary advantages of this method are; holistic view, actual physical representation of various electrical activities in a single nerve cell, display of the activity of all nerve cells flexibility with respect to network parameters. Moreover, this model can effectively help push forward our general ability to explore and conceptualize the electrical activity of interconnected networks of nerve cell behaving in concert. Also, this electronic module technique is the best of various means for this task of realistic representation of aggregates of neurons.gregates of neurons.

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Performance Analysis of Single Crystal Solar Cell by Impedance Measurement (임피던스 측정법을 이용한 단결정태양전지의 성능 특성 분석)

  • Jung, You-Ra;Choi, Yong-Sung;Hwang, Jong-Sun;Lee, Kyung-Sup
    • Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
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    • 2010.06a
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    • pp.202-202
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    • 2010
  • This study focused on the characteristics of single crystal solar cell using the impedance technique. In this experiment, the impedance was measured according to frequency's from 1mHz until 2MHz. The solar cell is R-L-C series circuit. Capacitance reactance was changed according to changing from low frequency to high frequency. It could know that the impedance was changed according to the frequency increases in solar cell.

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Fabrication and Characterization of Thermally Actuated Bimorph Probe for Living Cell Measurements with Experimental and Numerical Analysis

  • Cho Young-Hak;Kang Beom-Joon;Hong Seok-Kwan;Kang Jeong-Jin
    • Journal of Mechanical Science and Technology
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    • v.20 no.3
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    • pp.297-309
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    • 2006
  • This paper deals with a novel structure for single-cell characterization which makes use of bimorph micro thermal actuators combined with electrical sensor device and integrated microfluidic channel. The goal for this device is to capture and characterize individual biocell. Quantitative and qualitative characteristics of bimorph thermal actuator were analyzed with finite element analysis methods. Furthermore, optimization for the dimension of cantilevers and integrated parallel probe systems with microfluidic channels is able to be realized through the virtual simulation for actuation and the practical fabrication of prototype of probes. The experimental value of probe deflection was in accordance with the simulated one.

Design and Implementation of Flexible Sensor to Measure Mechanical Stiffness of Soft Particles (Soft Particle의 강성 측정을 위한 단순한 구조의 유연 물질 센서의 개발)

  • Ihn, Yong Seok;Yang, Minho;Koo, Ja Choon
    • The Journal of Korea Robotics Society
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    • v.11 no.3
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    • pp.133-139
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    • 2016
  • Increasing interest of human health, building bio-database (Bio DB) has been become a hot issue in life science. Consequently, Single Cell Analysis (SCA) which can explain biodiversity of lives has been a significant factor for building Bio DB. In numerous studies from these analyses, they have been showed that mechanical properties of cells can serve explanation of biological heterogeneity and criterion of disease states. Therefore, measuring mechanical properties of cells have great potential to be used in bio-medical applications. However, traditionally, many researchers have undergone difficult and time consuming work because handling small sized cells usually requires high-skilled technique. Thus, this paper shows robotized stiffness measurement technique using fixed ended beam sensor, precision motorized stage and substrate which have wall structure.

Affinity chromatography and capillary electrophoresis for analysis of the yeast ribosomal proteins

  • Goyder, Miriam S.;Willison, Keith R.;Klug, David R.;DeMello, Andrew J.;Ces, Oscar
    • BMB Reports
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    • v.45 no.4
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    • pp.233-238
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    • 2012
  • We present a top down separation platform for yeast ribosomal proteins using affinity chromatography and capillary electrophoresis which is designed to allow deposition of proteins onto a substrate. FLAG tagged ribosomes were affinity purified, and rRNA acid precipitation was performed on the ribosomes followed by capillary electrophoresis to separate the ribosomal proteins. Over 26 peaks were detected with excellent reproducibility (<0.5% RSD migration time). This is the first reported separation of eukaryotic ribosomal proteins using capillary electrophoresis. The two stages in this workflow, affinity chromatography and capillary electrophoresis, share the advantages that they are fast, flexible and have small sample requirements in comparison to more commonly used techniques. This method is a remarkably quick route from cell to separation that has the potential to be coupled to high throughput readout platforms for studies of the ribosomal proteome.

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.

Study on the Long-term Reliability of Solar Cell by High Temperature & Humidity Test (고온고습 시험을 통한 태양전지의 장기 신뢰성에 관한 연구)

  • Kang, Min-Soo;Jeon, Yu-Jae;Kim, Do-Seok;Shin, Young-Eui
    • Journal of Energy Engineering
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    • v.21 no.3
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    • pp.243-248
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    • 2012
  • In this study, The report analysed the characteristics of power drop and damage of surface in solar cell through high temperature and humidity test. The solar cells were tested during the 1000hr in $85^{\circ}C$ temperature and 85% humidity conditions, that excerpted standard of PV Module(KS C IEC-61215). An analysis of the cell surface through EL(Electroluminescence), the cell has partly change of surface in yearly. Single-crystalline Solar cell efficiency is decreased from 17.7% to 15.6% and decreasing rate is 11.9%. On the other hand, Poly-crystalline Solar cell efficiency is decreased from 15.5% to 14.0% and decreasing rate is 9.3%. A comparison of the fill factor for analysis of electro characteristic in yearly, Single-crystalline Solar cell efficiency is decreased from 78.7% to 78.1% and decreasing rate is 4.7%. On the other hand, Poly-crystalline Solar cell efficiency is decreased from 78.1% to 76.7% and decreasing rate is 1.8%. Single-crystalline has more bigger power drop than poly-crystalline by the silicon purity and silicon atom arrangement. Also, FF decreasing rate has more bigger drop than efficiency decreasing rate for the reason that the damage of surface by exterior environmental factor is the more influence in cell than other reason that is decreasing FF by damage of p-n junction.