Development and characterization of five novel cell lines from snubnose pompano, Trachinotus blochii (Lacepede, 1801), and their application in gene expression and virological studies

2021 ◽  
Author(s):  
Nafeez Ahmed Abdul ◽  
Abdul Majeed Seepoo ◽  
Taju Gani ◽  
Vimal Sugumar ◽  
Suryakodi Selvam ◽  
...  
Keyword(s):  
Gene Expression ◽  
Cell Lines ◽  
Trachinotus Blochii

BIOM-45. CHARACTERIZATION OF EVs RELEASED FROM 5-ALA DOSED GLIAL AND EXTRA-AXIAL TUMORS

2021 ◽  
Vol 23 (Supplement_6) ◽  
pp. vi21-vi21
Author(s):  
Tiffaney Hsia ◽  
Anudeep Yekula ◽  
Bob Carter ◽  
Leonora Balaj
Keyword(s):  
Gene Expression ◽  
Tumor Cells ◽  
Cell Lines ◽  
Fluorescence Analysis ◽  
Illumina Miseq ◽  
Biological Regulation ◽  
Guided Surgery ◽  
Antibody Staining ◽  
Fluorescence Guided Surgery

Abstract INTRODUCTION The administration of 5-Aminolevunlinic acid (5-ALA) in the context of fluorescence-guided surgery is contingent on the highly selective accumulation of PpIX in tumor cells. PpIX is a precursor in the heme pathway that naturally exhibits fluorescent and phototoxic properties upon excitation. We have previously reported evidence of PpIX fluorescence in tumor-specific extracellular vesicles (EVs). Here, we explore the implications of exogenous 5-ALA on cellular function, EV characteristics, and gene expression. METHODS We have characterized a range of glioblastoma, meningioma, and healthy cell lines (Gli36 EGFRvIII, Gli36 EGFR WT, U87, CH-157, IOMM-Lee, and HBMVEC). At equal confluency, cells were dosed with 0.8 mM 5-ALA or mock dosed. Media was collected and processed to eliminate debris after 24 hours. Analysis of cells and EVs was conducted using Amnis ImageStream (IFC) and Nanoparticle Tracking Analysis. EV subpopulations were analyzed with IFC following antibody staining. RNA was extracted using Qiagen exoRNeasy and RNeasy. Libraries were prepared via Qiagen UPX 3’ Transcriptome kit and sequenced using Illumina MiSeq. Data analysis was carried out via GeneGlobe, CLC workbench, and MATLAB. RESULTS Following 5-ALA dosing, all tumor cells and their derived EVs exhibit significant levels of fluorescence. Analysis of EV subpopulations demonstrated a general decrease in tetraspanin-positive EVs following 5-ALA dosing. At 24 hours, Gli36 cell lines exhibited increased EV release post 5-ALA whereas U87 and meningioma lines resulted in a decreased EV release rate. This clustering is also reflected in EV size distributions and in cellular and EV differential gene expression analysis. Gene ontology analysis of common genes from mock and dosed EVs demonstrated high counts of genes controlling biological regulation as well as cellular and metabolic processes. CONCLUSION Collection and characterization of cancer specific EVs may be advantageous to liquid biopsy development.

Characterization of newly established clonal oviductal cell lines and differential hormonal reculation of gene expression

2003 ◽  
Vol 39 (3-4) ◽  
pp. 146-156 ◽  
Author(s):  
Tomohiro Umezu ◽  
Makoto Hanazono ◽  
Shinichi Aizawa ◽  
Yasuhiro Tomooka
Keyword(s):  
Gene Expression ◽  
Cell Lines ◽  
Oviductal Cell

scholarly journals Gene Expression Music Algorithm-Based Characterization of the Ewing Sarcoma Stem Cell Signature

2016 ◽  
Vol 2016 ◽  
pp. 1-10 ◽  
Author(s):  
Martin Sebastian Staege
Keyword(s):  
Gene Expression ◽  
Stem Cells ◽  
Stem Cell ◽  
Cell Lines ◽  
Microarray Data ◽  
Ewing Sarcoma ◽  
Correlation Coefficients ◽  
Music Algorithm ◽  
Dna Microarray Data

Gene Expression Music Algorithm (GEMusicA) is a method for the transformation of DNA microarray data into melodies that can be used for the characterization of differentially expressed genes. Using this method we compared gene expression profiles from endothelial cells (EC), hematopoietic stem cells, neuronal stem cells, embryonic stem cells (ESC), and mesenchymal stem cells (MSC) and defined a set of genes that can discriminate between the different stem cell types. We analyzed the behavior of public microarray data sets from Ewing sarcoma (“Ewing family tumors,” EFT) cell lines and biopsies in GEMusicA after prefiltering DNA microarray data for the probe sets from the stem cell signature. Our results demonstrate that individual Ewing sarcoma cell lines have a high similarity to ESC or EC. Ewing sarcoma cell lines with inhibited Ewing sarcoma breakpoint region 1-Friend leukemia virus integration 1 (EWSR1-FLI1) oncogene retained the similarity to ESC and EC. However, correlation coefficients between GEMusicA-processed expression data between EFT and ESC decreased whereas correlation coefficients between EFT and EC as well as between EFT and MSC increased after knockdown of EWSR1-FLI1. Our data support the concept of EFT being derived from cells with features of embryonic and endothelial cells.

Development and characterization of novel cell lines from Etroplus suratensis and their applications in virology, toxicology and gene expression

2012 ◽  
Vol 80 (2) ◽  
pp. 312-334 ◽  
Author(s):  
V. Sarath Babu ◽  
V. Chandra ◽  
K. S. N. Nambi ◽  
S. A. Majeed ◽  
G. Taju ◽  
...  
Keyword(s):  
Gene Expression ◽  
Cell Lines ◽  
Etroplus Suratensis

scholarly journals Isolation and Characterization of the cDNA for Mouse Neutrophil Collagenase: Demonstration of Shared Negative Regulatory Pathways for Neutrophil Secondary Granule Protein Gene Expression

Blood ◽  
1998 ◽  
Vol 91 (7) ◽  
pp. 2517-2524 ◽  
Author(s):  
Nathan D. Lawson ◽  
Arati Khanna-Gupta ◽  
Nancy Berliner
Keyword(s):  
Gene Expression ◽  
Cell Line ◽  
Cell Lines ◽  
Regulatory Pathways ◽  
Granule Protein ◽  
Isolation And Characterization ◽  
Negative Regulatory Pathway ◽  
Neutrophil Collagenase ◽  
Secondary Granule

A characteristic of normal neutrophil maturation is the induction of secondary granule protein (SGP) mRNA expression. Several leukemic human cell lines mimic normal morphologic neutrophil differentiation but fail to express SGPs, such as lactoferrin (LF) and neutrophil gelatinase (NG). In contrast, two murine cell lines (32D C13 and MPRO) are able to differentiate into neutrophils and induce expression of LF and NG. Therefore, to study the normal regulation and function of these genes, the corresponding murine homologs must be isolated. Using cDNA representational difference analysis (RDA) to compare a committed myeloid progenitor cell line (EPRO) with the multipotent stem cell line from which it was derived (EML), we isolated a fragment bearing homology to human neutrophil collagenase (hNC). Here, we describe the cloning and characterization of a full-length (∼2 kb) clone that exhibits nearly 65% nucleotide and 73% amino acid identity to hNC. Ribonuclease protection analysis (RPA) of the tissues and cell lines shows that mouse NC (mNC) is expressed only in cell lines exhibiting neutrophilic characteristics, further confirming its identity as the mouse homolog of hNC. Furthermore, we have demonstrated a shared negative regulatory pathway for this and other SGP genes. We have previously shown that CCAAT displacement protein (CDP/cut) binds to a specific region of the LF promoter, and overexpression of CDP blocks G-CSF–induced upregulation of LF gene expression in 32D C13 cells. We show here that in these cells, upregulation of both NC and NG is also blocked. CDP is thus the first identified transcription factor that is a candidate for mediating the shared regulation of neutrophil SGP protein genes.

Establishment and Characterization of Gene Expression of Two New Ovine Mammary Cell Lines: Preliminary Results.

2011 ◽  
Vol 85 (Suppl_1) ◽  
pp. 350-350
Author(s):  
Mariana Ianello Giassetti ◽  
Camilla Mota Mendes ◽  
Flavia Regina Oliveira de Barros ◽  
Mayra Elena Ortiz D'Avila Assumpcao ◽  
Jose Antonio Visintin
Keyword(s):  
Gene Expression ◽  
Cell Lines ◽  
Preliminary Results ◽  
Mammary Cell

Array-CGH and Gene Expression Profiling Based Molecular Characterization of Myeloid Leukemia Cell Lines.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 4397-4397
Author(s):  
Frank G. Rucker ◽  
Sandrine Sander ◽  
Konstanze Dohner ◽  
Hartmut Dohner ◽  
Jonathan R. Pollack ◽  
...  
Keyword(s):  
Gene Expression ◽  
Molecular Characterization ◽  
Cell Lines ◽  
Myeloid Leukemia ◽  
Leukemia Cell ◽  
Microarray Platform ◽  
Model Systems ◽  
Cytogenetic Aberrations ◽  
Leukemia Cell Lines

Abstract Myeloid leukemias are characterized by cytogenetic and molecular-genetic aberrations resulting in altered gene expression. Nevertheless, so far still little is known regarding the underlying mechanisms of leukemogenesis. To model and investigate the different aspects of leukemia pathogenesis a widely accepted approach is to use immortalized leukemia cell lines. While these provide powerful tools for the investigation of gene function, the dissection of signal transduction pathways and the analysis of drug effects, to our knowledge only few studies have addressed the question whether hematopoietic cell lines represent reliable model systems. In order to improve the molecular characterization of these model systems we therefore analyzed 18 myeloid leukemia cell lines using DNA microarray technology. To determine the secondary aberrations acquired in addition to their characteristic primary cytogenetic aberrations during numerous passages in vitro, we first analyzed all cell lines by array-CGH (comparative genomic hybridization). Using a BAC/PAC platform with an average resolution of ~ 1.5 Mb across the entire genome we identified recurrent losses and gains, as well as high level amplifications like e.g. an amplification in 4q12 (Kasumi1) and in 8q24 (HL60). The parallel analysis of gene expression using a whole genome cDNA microarray platform helped to further delineate potential candidate genes in the affected regions (e.g. overexpression of KIT in the 4q12 and MYC in the 8q24 amplicon). Furthermore, unsupervised hierarchical cluster analysis revealed distinct gene signatures pointing towards dysregulated transcriptional networks. Comparison of our findings with acute myeloid leukemia patient data (Bullinger et al. 2004) showed the signatures underlying characteristic cytogenetic aberrations like e.g. t(15;17) were conserved in cell lines. Interestingly, these signatures were also quite robust as they displayed a highly significant correlation with published cell line data profiled on a different DNA microarray platform in a different laboratory. Thus, our analyses demonstrate that cell lines exhibit conserved signatures correlating with the primary balanced cytogenetic aberrations, and that most cell lines even when grown and analyzed under different conditions provide highly robust signatures. Therefore, our refined molecular characterization of myeloid cell lines supports the utility of cell lines as powerful model systems and provides additional insights into the molecular mechanisms of leukemogenesis.

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