scholarly journals Extracellular vesicle and particle isolation from human and murine cell lines, tissues, and bodily fluids

2020 ◽  
pp. 100225
Author(s):  
Linda Bojmar ◽  
Han Sang Kim ◽  
Gabriel C. Tobias ◽  
Fanny A. Pelissier Vatter ◽  
Serena Lucotti ◽  
...  
Keyword(s):  
Cell Lines ◽  
Extracellular Vesicle ◽  
Murine Cell ◽  
Bodily Fluids

scholarly journals Differences in Extracellular Vesicle Protein Cargo Are Dependent on Head and Neck Squamous Cell Carcinoma Cell of Origin and Human Papillomavirus Status

Cancers ◽  
2021 ◽  
Vol 13 (15) ◽  
pp. 3714
Author(s):  
Christine Goudsmit ◽  
Felipe da Veiga Leprevost ◽  
Venkatesha Basrur ◽  
Lila Peters ◽  
Alexey Nesvizhskii ◽  
...  
Keyword(s):  
Squamous Cell Carcinoma ◽  
Cell Carcinoma ◽  
Head And Neck ◽  
Cell Lines ◽  
Squamous Cell ◽  
Extracellular Vesicle ◽  
Neck Squamous Cell Carcinoma ◽  
Cytokeratin 19 ◽  
Oral Keratinocytes ◽  
Transformed Cell Lines

To identify potential extracellular vesicle (EV) biomarkers in head and neck squamous cell carcinoma (HNSCC), we evaluated EV protein cargo and whole cell lysates (WCL) from HPV-positive and -negative HNSCC cell lines, as well as normal oral keratinocytes and HPV16-transformed cells. EVs were isolated from serum-depleted, conditioned cell culture media by polyethylene glycol (PEG) precipitation/ultracentrifugation. EV and WCL preparations were analyzed by LC-MS/MS. Candidate proteins detected at significantly higher levels in EV compared with WCL, or compared with EV from normal oral keratinocytes, were identified and confirmed by Wes Simple Western protein analysis. Our findings suggest that these proteins may be potential HNSCC EV markers as proteins that may be (1) selectively included in EV cargo for export from the cell as a strategy for metastasis, tumor cell survival, or modification of tumor microenvironment, or (2) representative of originating cell composition, which may be developed for diagnostic or prognostic use in clinical liquid biopsy applications. This work demonstrates that our method can be used to reliably detect EV proteins from HNSCC, normal keratinocyte, and transformed cell lines. Furthermore, this work has identified HNSCC EV protein candidates for continued evaluation, specifically tenascin-C, HLA-A, E-cadherin, EGFR, EPHA2, and cytokeratin 19.

Advances in Microfluidic Extracellular Vesicle Analysis for Cancer Diagnostics

Lab on a Chip ◽  
2021 ◽  
Author(s):  
Shibo Cheng ◽  
Yutao Li ◽  
He Yan ◽  
Yunjie Wen ◽  
Xin Zhou ◽  
...  
Keyword(s):  
Extracellular Vesicles ◽  
Extracellular Vesicle ◽  
Cancer Diagnostics ◽  
Bodily Fluids ◽  
Intercellular Communications

Extracellular vesicles (EVs) secreted by cells into the bloodstream and other bodily fluids, including exosomes, have been demonstrated to be a class of significant messengers that mediate intercellular communications. Tumor-derived...

scholarly journals Transfection of the genes for interleukin-12 into the K1735 melanoma and the EMT6 mammary sarcoma murine cell lines reveals distinct mechanisms of antitumor activity

2003 ◽  
Vol 106 (5) ◽  
pp. 690-698 ◽  
Author(s):  
James P. Moran ◽  
Scott A. Gerber ◽  
Celeste A. Martin ◽  
John G. Frelinger ◽  
Edith M. Lord
Keyword(s):  
Antitumor Activity ◽  
Cell Lines ◽  
Interleukin 12 ◽  
Murine Cell

scholarly journals Design and synthesis of novel 1,3,5-triphenyl pyrazolines as potential anti-inflammatory agents through allosteric inhibition of protein kinase Czeta (PKCζ)

MedChemComm ◽  
2018 ◽  
Vol 9 (6) ◽  
pp. 1076-1082 ◽  
Author(s):  
Mohammad Abdel-Halim ◽  
Ashraf H. Abadi ◽  
Matthias Engel
Keyword(s):  
Protein Kinase ◽  
Mouse Models ◽  
Cell Lines ◽  
Lead Compound ◽  
Allosteric Inhibition ◽  
Murine Cell ◽  
Design And Synthesis ◽  
Anti Inflammatory

A new focused library of PKCζ inhibitors was synthesized, leading to the identification of compound2h. Owing to its improved cellular potency in human and murine cell lines, this new lead compound opens up the possibility to evaluate allosteric PKCζ inhibitors in rat or mouse models.

scholarly journals A proteomic approach to understand the clinical significance of acute myeloid leukemia-derived extracellular vesicles reflecting essential characteristics of leukemia

2020 ◽  
pp. mcp.RA120.002169
Author(s):  
Ka-Won Kang ◽  
Hyoseon Kim ◽  
Woojune Hur ◽  
Jik-han Jung ◽  
Su Jin Jeong ◽  
...  
Keyword(s):  
Acute Myeloid Leukemia ◽  
Cell Lines ◽  
Myeloid Leukemia ◽  
Extracellular Vesicle ◽  
The Cancer Genome Atlas ◽  
Enzyme Linked Immunosorbent Assay ◽  
Expression Levels ◽  
Proteomic Approach ◽  
Cancer Genome Atlas ◽  
Acute Myeloid

Extracellular vesicle (EV) proteins from acute myeloid leukemia (AML) cell lines were analyzed using mass spectrometry. The analyses identified 2450 proteins, including 461 differentially expressed proteins (290 upregulated and 171 downregulated). CD53 and CD47 were upregulated and were selected as candidate biomarkers. The association between survival of patients with AML and the expression levels of CD53 and CD47 at diagnosis was analyzed using mRNA expression data from The Cancer Genome Atlas database. Patients with higher expression levels showed significantly inferior survival than those with lower expression levels. Enzyme-linked immunosorbent assay results of the expression levels of CD53 and CD47 from EVs in the bone marrow of patients with AML at diagnosis and at the time of complete remission with induction chemotherapy revealed that patients with downregulated CD53 and CD47 expression appeared to relapse less frequently. Network model analysis of EV proteins revealed several upregulated kinases, including LYN, CSNK2A1, SYK, CSK, and PTK2B. The potential cytotoxicity of several clinically applicable drugs that inhibit these kinases was tested in AML cell lines. The drugs lowered the viability of AML cells. The collective data suggest that AML-derived EVs could reflect essential leukemia biology.

scholarly journals Syncytin 1 dependent horizontal transfer of marker genes from retrovirally transduced cells

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Berna Uygur ◽  
Kamran Melikov ◽  
Anush Arakelyan ◽  
Leonid B. Margolis ◽  
Leonid V. Chernomordik
Keyword(s):  
Cell Lines ◽  
Membrane Vesicles ◽  
Retroviral Vectors ◽  
Extracellular Vesicle ◽  
Marker Genes ◽  
Target Cells ◽  
Retroviral Transduction ◽  
Gfp Gene ◽  
Safety Test

AbstractRetroviral transduction is routinely used to generate cell lines expressing exogenous non-viral genes. Here, we show that human cells transduced to stably express GFP transfer GFP gene to non-transduced cells. This horizontal gene transfer was mediated by a fraction of extracellular membrane vesicles that were released by the transduced cells. These vesicles carried endogenous retroviral envelope protein syncytin 1 and essentially acted as replication-competent retroviruses. The ability to transfer the GFP gene correlated with the levels of syncytin 1 expression in the transduced cells and depended on the fusogenic activity of this protein, substantiating the hypothesis that endogenous syncytin 1 mediates fusion stage in the delivery of extracellular vesicle cargo into target cells. Our findings suggest that testing for replication-competent retroviruses, a routine safety test for transduced cell products in clinical studies, should be also carried out for cell lines generated by retroviral vectors in in vitro studies.

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