scholarly journals Analysis of SNARE-Mediated Exocytosis Using a Cell Fusion Assay

10.5772/29580 ◽  
2012 ◽  
Author(s):  
Chuan Hu ◽  
Nazarul Hasan ◽  
Krista Riggs
Keyword(s):  
Cell Fusion ◽  
A Cell

Quantitative cell fusion: the fusion sensitivity (FS) potential

1981 ◽  
Vol 49 (1) ◽  
pp. 87-97
Author(s):  
D. Rohme
Keyword(s):  
Cell Line ◽  
Cell Fusion ◽  
Cell Lines ◽  
Dose Response ◽  
Sendai Virus ◽  
Biological Significance ◽  
Diploid Cell ◽  
Mammalian Cell Lines ◽  
Virus Dose ◽  
A Cell

The dose response of Sendai virus-induced cell fusion was studied in 10 mammalian cell lines, comprising 5 continuous and 5 diploid cell lines originating from 5 species. The extent of fusion was calculated using a parameter directly proportional to the number of fusion events (t-parameter). At lower levels of fusion the dose response was found to be based on the same simple kinetic rules in all cell lines and was defined by the formula: t = FS. FAU/(I + FS. FAU), where FS (fusion sensitivity) is a cell-specific constant of the fusion rate and FAU (fusion activity units) is the virus dose. The FS potential of a cell line was determined as the linear regression coefficient of the fusion index (t/(I - t)) on the virus dose. At higher levels of fusion, when the fusion extent reached cell-line-specific maximal levels, the dose response was not as uniform. In general, and particularly in the cases of the diploid cell lines, these maximal levels were directly proportional to the FS potentials. Thus, it was concluded that the FS potential is the basic quantitative feature, which expresses the cellular fusion efficiency. The fact that FS varied extensively between cell lines, but at the same time apparently followed certain patterns (being higher in continuous compared to diploid cell lines and being related to the species of origin of the cells), emphasizes it biological significance as well as its possible usefulness in studies of the efficiency of various molecular interactions in the cell membrane/cytoskeleton system.

Estimation of the number of monoclonal hybridomas in a cell fusion experiment. Effect of post-fusion cell dilution on hybridoma survival

1981 ◽  
Vol 45 (2) ◽  
pp. 109-115 ◽  
Author(s):  
Angel L. De Blas ◽  
Makarand V. Ratnaparkhi ◽  
James E. Mosimann
Keyword(s):  
Cell Fusion ◽  
A Cell ◽  
Fusion Cell

[4] Estimation of the number of monoclonal hybridomas in a cell-fusion experiment

1983 ◽  
pp. 36-39 ◽  
Author(s):  
Angel L. De Blas ◽  
Makarand V. Ratnaparkhi ◽  
James E. Mosimann
Keyword(s):  
Cell Fusion ◽  
A Cell

Rapid screening method with a cell multisizer for inhibitors of human immunodeficiency virus-induced cell fusion in vitro.

1988 ◽  
Vol 26 (6) ◽  
pp. 1229-1232 ◽  
Author(s):  
H Nakashima ◽  
A Tanabe ◽  
T S Tochikura ◽  
N Yamamoto
Keyword(s):  
Human Immunodeficiency Virus ◽  
Cell Fusion ◽  
Screening Method ◽  
Rapid Screening ◽  
Immunodeficiency Virus ◽  
A Cell ◽  
Rapid Screening Method

scholarly journals The Anticoagulant Nafamostat Potently Inhibits SARS-CoV-2 S Protein-Mediated Fusion in a Cell Fusion Assay System and Viral Infection In Vitro in a Cell-Type-Dependent Manner

Viruses ◽  
2020 ◽  
Vol 12 (6) ◽  
pp. 629 ◽  
Author(s):  
Mizuki Yamamoto ◽  
Maki Kiso ◽  
Yuko Sakai-Tagawa ◽  
Kiyoko Iwatsuki-Horimoto ◽  
Masaki Imai ◽  
...  
Keyword(s):  
Cell Fusion ◽  
Assay System ◽  
Dependent Manner ◽  
Cell Type ◽  
S Protein ◽  
Candidate Drug ◽  
A Cell ◽  
Transmembrane Serine Protease ◽  
Nafamostat Mesylate

Although infection by SARS-CoV-2, the causative agent of coronavirus pneumonia disease (COVID-19), is spreading rapidly worldwide, no drug has been shown to be sufficiently effective for treating COVID-19. We previously found that nafamostat mesylate, an existing drug used for disseminated intravascular coagulation (DIC), effectively blocked Middle East respiratory syndrome coronavirus (MERS-CoV) S protein-mediated cell fusion by targeting transmembrane serine protease 2 (TMPRSS2), and inhibited MERS-CoV infection of human lung epithelium-derived Calu-3 cells. Here we established a quantitative fusion assay dependent on severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) S protein, angiotensin I converting enzyme 2 (ACE2) and TMPRSS2, and found that nafamostat mesylate potently inhibited the fusion while camostat mesylate was about 10-fold less active. Furthermore, nafamostat mesylate blocked SARS-CoV-2 infection of Calu-3 cells with an effective concentration (EC)50 around 10 nM, which is below its average blood concentration after intravenous administration through continuous infusion. On the other hand, a significantly higher dose (EC50 around 30 μM) was required for VeroE6/TMPRSS2 cells, where the TMPRSS2-independent but cathepsin-dependent endosomal infection pathway likely predominates. Together, our study shows that nafamostat mesylate potently inhibits SARS-CoV-2 S protein-mediated fusion in a cell fusion assay system and also inhibits SARS-CoV-2 infection in vitro in a cell-type-dependent manner. These findings, together with accumulated clinical data regarding nafamostat’s safety, make it a likely candidate drug to treat COVID-19.

scholarly journals Usefulness of a Non-invasive Reporter System for Monitoring Reprogramming State in Pig Cells: Results of a Cell Fusion Experiment

2010 ◽  
Vol 56 (4) ◽  
pp. 363-369 ◽  
Author(s):  
Akio OZAWA ◽  
Eri AKASAKA ◽  
Satoshi WATANABE ◽  
Mitsutoshi YOSHIDA ◽  
Kazuchika MIYOSHI ◽  
...  
Keyword(s):  
Cell Fusion ◽  
Reporter System ◽  
Non Invasive ◽  
A Cell

scholarly journals Targeted disruption of genes for gp138, a cell-fusion-related protein in Dictyostelium discoideum, revealed the existence of a third gene

1996 ◽  
Vol 38 (3) ◽  
pp. 271-279 ◽  
Author(s):  
Nobuyuki Yamaguchi ◽  
Mikihiko Higa ◽  
Kazuhiro Aiba ◽  
Hui Fang ◽  
Yoshimasa Tanaka ◽  
...  
Keyword(s):  
Dictyostelium Discoideum ◽  
Cell Fusion ◽  
Related Protein ◽  
Targeted Disruption ◽  
A Cell

scholarly journals Human cord blood CD34+ progenitor cells acquire functional cardiac properties through a cell fusion process

2011 ◽  
Vol 300 (5) ◽  
pp. H1875-H1884 ◽  
Author(s):  
Daniele Avitabile ◽  
Alessia Crespi ◽  
Chiara Brioschi ◽  
Valeria Parente ◽  
Gabriele Toietta ◽  
...  
Keyword(s):  
Cord Blood ◽  
Cell Fusion ◽  
Fluorescent Protein ◽  
Lentiviral Vector ◽  
Fusion Process ◽  
Resting Potential ◽  
Human Cord Blood ◽  
Neonatal Cardiomyocytes ◽  
Cord Blood Cd34 ◽  
A Cell

The efficacy of cardiac repair by stem cell administration relies on a successful functional integration of injected cells into the host myocardium. Safety concerns have been raised about the possibility that stem cells may induce foci of arrhythmia in the ischemic myocardium. In a previous work ( 36 ), we showed that human cord blood CD34+ cells, when cocultured on neonatal mouse cardiomyocytes, exhibit excitation-contraction coupling features similar to those of cardiomyocytes, even though no human genes were upregulated. The aims of the present work are to investigate whether human CD34+ cells, isolated after 1 wk of coculture with neonatal ventricular myocytes, possess molecular and functional properties of cardiomyocytes and to discriminate, using a reporter gene system, whether cardiac differentiation derives from a (trans)differentiation or a cell fusion process. Umbilical cord blood CD34+ cells were isolated by a magnetic cell sorting method, transduced with a lentiviral vector carrying the enhanced green fluorescent protein (EGFP) gene, and seeded onto primary cultures of spontaneously beating rat neonatal cardiomyocytes. Cocultured EGFP+/CD34+-derived cells were analyzed for their electrophysiological features at different time points. After 1 wk in coculture, EGFP+ cells, in contact with cardiomyocytes, were spontaneously contracting and had a maximum diastolic potential (MDP) of −53.1 mV, while those that remained isolated from the surrounding myocytes did not contract and had a depolarized resting potential of −11.4 mV. Cells were then resuspended and cultured at low density to identify EGFP+ progenitor cell derivatives. Under these conditions, we observed single EGFP+ beating cells that had acquired an hyperpolarization-activated current typical of neonatal cardiomyocytes (EGFP+ cells, −2.24 ± 0.89 pA/pF; myocytes, −1.99 ± 0.63 pA/pF, at −125 mV). To discriminate between cell autonomous differentiation and fusion, EGFP+/CD34+ cells were cocultured with cardiac myocytes infected with a red fluorescence protein-lentiviral vector; under these conditions we found that 100% of EGFP+ cells were also red fluorescent protein positive, suggesting cell fusion as the mechanism by which cardiac functional features are acquired.

scholarly journals Persistent Infection of a Cell Line of Mouse Origin after Cell Fusion by u.v.-inactivated Sendai Virus

1980 ◽  
Vol 46 (1) ◽  
pp. 219-226 ◽  
Author(s):  
T. J. Schnitzer ◽  
J. F. Watkins
Keyword(s):  
Cell Line ◽  
Cell Fusion ◽  
Persistent Infection ◽  
Sendai Virus ◽  
A Cell

Cell fusion, using sendai virus, to effect inter-species transfer of a cell-associated parasite (Theileria parva)

1974 ◽  
Vol 4 (5) ◽  
pp. 519-521 ◽  
Author(s):  
A IRVIN ◽  
C BROWN ◽  
G KANHAI ◽  
D STAGG ◽  
L ROWE
Keyword(s):  
Cell Fusion ◽  
Sendai Virus ◽  
Theileria Parva ◽  
A Cell
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