High-Throughput Identification of Combinatorial Ligands for DNA Delivery in Cell Culture

2008 ◽  
Author(s):  
Mathias G. Svahn ◽  
Kersten S. Rabe ◽  
Geoffrey Barger ◽  
Samir EL-Andaloussi ◽  
Oscar E. Simonson ◽  
...  
Keyword(s):  
Cell Culture ◽  
High Throughput ◽  
Dna Delivery

scholarly journals Real-time imaging of cellular forces using optical interference

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Andrew T. Meek ◽  
Nils M. Kronenberg ◽  
Andrew Morton ◽  
Philipp Liehm ◽  
Jan Murawski ◽  
...  
Keyword(s):  
Cell Culture ◽  
Real Time ◽  
High Throughput ◽  
Mechanical Activity ◽  
Dynamic Processes ◽  
Imaging Method ◽  
Neonatal Cardiomyocytes ◽  
A Cell ◽  
Cellular Forces ◽  
Time Acquisition

AbstractImportant dynamic processes in mechanobiology remain elusive due to a lack of tools to image the small cellular forces at play with sufficient speed and throughput. Here, we introduce a fast, interference-based force imaging method that uses the illumination of an elastic deformable microcavity with two rapidly alternating wavelengths to map forces. We show real-time acquisition and processing of data, obtain images of mechanical activity while scanning across a cell culture, and investigate sub-second fluctuations of the piconewton forces exerted by macrophage podosomes. We also demonstrate force imaging of beating neonatal cardiomyocytes at 100 fps which reveals mechanical aspects of spontaneous oscillatory contraction waves in between the main contraction cycles. These examples illustrate the wider potential of our technique for monitoring cellular forces with high throughput and excellent temporal resolution.

scholarly journals Development of NS3/4A Protease-Based Reporter Assay Suitable for Efficiently Assessing Hepatitis C Virus Infection

2009 ◽  
Vol 53 (11) ◽  
pp. 4825-4834 ◽  
Author(s):  
Kao-Lu Pan ◽  
Jin-Ching Lee ◽  
Hsing-Wen Sung ◽  
Teng-Yuang Chang ◽  
John T.-A. Hsu
Keyword(s):  
Cell Culture ◽  
Life Cycle ◽  
Hepatitis C Virus ◽  
Hepatitis C ◽  
Virus Infection ◽  
High Throughput ◽  
High Throughput Screening ◽  
Reporter System ◽  
Viral Protease ◽  
Peptide Cleavage

ABSTRACT A cell culture system for the production of hepatitis C virus (HCV) whole virions has greatly accelerated studies of the virus life cycle and the discovery of anti-HCV agents. However, the quantification of the HCV titers in a whole-virus infection/replication system currently relies mostly on reverse transcription-PCR or immunofluorescence assay, which would be cumbersome for high-throughput drug screening. To overcome this problem, this study has generated a novel cell line, Huh7.5-EG(Δ4B5A)SEAP, that carries a dual reporter, EG(Δ4B5A)SEAP. The EG(Δ4B5A)SEAP reporter is a viral protease-cleavable fusion protein in which the enhanced green fluorescence protein is linked to secreted alkaline phosphatase (SEAP) in frame via Δ4B5A, a short peptide cleavage substrate for NS3/4A viral protease. This study demonstrates that virus replication/infection in the Huh7.5-EG(Δ4B5A)SEAP cells can be quantitatively indicated by measuring the SEAP activity in cell culture medium. The levels of SEAP released from HCV-infected Huh7.5-EG(Δ4B5A)SEAP cells correlated closely with the amounts of HCV in the inocula. The Huh7.5-EG(Δ4B5A)SEAP cells were also shown to be a suitable host for the discovery of anti-HCV inhibitors by using known compounds that target multiple stages of the HCV life cycle. The Z′-factor of this assay ranged from 0.64 to 0.74 in 96-well plates, indicating that this reporter system is suitable for high-throughput screening of prospective anti-HCV agents.

A high-throughput cell culture system based on capillary and centrifugal actions for rapid antimicrobial susceptibility testing

Lab on a Chip ◽  
2020 ◽  
Vol 20 (24) ◽  
pp. 4552-4560
Author(s):  
Taegeun Lim ◽  
Eun-Geun Kim ◽  
Jungil Choi ◽  
Sunghoon Kwon
Keyword(s):  
Cell Culture ◽  
High Throughput ◽  
Antimicrobial Susceptibility ◽  
Culture System ◽  
Susceptibility Testing ◽  
Culture Media ◽  
Antimicrobial Susceptibility Testing ◽  
Testing System ◽  
Cell Culture System

A capillary and centrifuge-based rapid antimicrobial susceptibility testing system is developed to reduce the time of loading the sample and culture media while achieving a high-throughput testing capacity.

scholarly journals Chemically diverse polymer microarrays and high throughput surface characterisation: a method for discovery of materials for stem cell culture

2014 ◽  
Vol 2 (11) ◽  
pp. 1604-1611 ◽  
Author(s):  
A. D. Celiz ◽  
J. G. W. Smith ◽  
A. K. Patel ◽  
R. Langer ◽  
D. G. Anderson ◽  
...  
Keyword(s):  
Cell Culture ◽  
Stem Cell ◽  
High Throughput ◽  
Screening Tool ◽  
New Materials ◽  
Surface Characterisation ◽  
Stem Cell Culture

Chemically diverse polymer microarrays as a powerful screening tool for the discovery of new materials for a variety of applications.

Patterned superhydrophobic surfaces to process and characterize biomaterials and 3D cell culture

2018 ◽  
Vol 5 (3) ◽  
pp. 379-393 ◽  
Author(s):  
A. I. Neto ◽  
P. A. Levkin ◽  
J. F. Mano
Keyword(s):  
Cell Culture ◽  
High Throughput ◽  
High Throughput Screening ◽  
3D Cell Culture ◽  
Superhydrophobic Surfaces ◽  
Technological Breakthrough ◽  
Large Numbers

Microarrays are a technological breakthrough for high-throughput screening of large numbers of assays.

scholarly journals High-Throughput Assessment of Mechanistic Toxicity of Chemicals in Miniaturized 3D Cell Culture

2018 ◽  
Vol 79 (1) ◽  
pp. e66
Author(s):  
Pranav Joshi ◽  
Soo-Yeon Kang ◽  
Akshata Datar ◽  
Moo-Yeal Lee
Keyword(s):  
Cell Culture ◽  
High Throughput ◽  
3D Cell Culture

scholarly journals High-Throughput Nano-Biofilm Microarray for Antifungal Drug Discovery

mBio ◽  
2013 ◽  
Vol 4 (4) ◽  
Author(s):  
Anand Srinivasan ◽  
Kai P. Leung ◽  
Jose L. Lopez-Ribot ◽  
Anand K. Ramasubramanian
Keyword(s):  
Cell Culture ◽  
Candida Albicans ◽  
Drug Discovery ◽  
High Throughput ◽  
Fungal Pathogen ◽  
Microarray Platform ◽  
Microbial Cell ◽  
Content Type ◽  
Petri Dishes ◽  
Opportunistic Fungal Pathogen

ABSTRACT Micro- and nanoscale technologies have radically transformed biological research from genomics to tissue engineering, with the relative exception of microbial cell culture, which is still largely performed in microtiter plates and petri dishes. Here, we present nanoscale culture of the opportunistic fungal pathogen Candida albicans on a microarray platform. The microarray consists of 1,200 individual cultures of 30 nl of C. albicans biofilms (“nano-biofilms”) encapsulated in an inert alginate matrix. We demonstrate that these nano-biofilms are similar to conventional macroscopic biofilms in their morphological, architectural, growth, and phenotypic characteristics. We also demonstrate that the nano-biofilm microarray is a robust and efficient tool for accelerating the drug discovery process: (i) combinatorial screening against a collection of 28 antifungal compounds in the presence of immunosuppressant FK506 (tacrolimus) identified six drugs that showed synergistic antifungal activity, and (ii) screening against the NCI challenge set small-molecule library identified three heretofore-unknown hits. This cell-based microarray platform allows for miniaturization of microbial cell culture and is fully compatible with other high-throughput screening technologies. IMPORTANCE Microorganisms are typically still grown in petri dishes, test tubes, and Erlenmeyer flasks in spite of the latest advances in miniaturization that have benefitted other allied research fields, including genomics and proteomics. Culturing microorganisms in small scale can be particularly valuable in cutting down time, cost, and reagent usage. This paper describes the development, characterization, and application of nanoscale culture of an opportunistic fungal pathogen, Candida albicans. Despite a more than 2,000-fold reduction in volume, the growth characteristics and drug response profiles obtained from the nanoscale cultures were comparable to the industry standards. The platform also enabled rapid identification of new drug candidates that were effective against C. albicans biofilms, which are a major cause of mortality in hospital-acquired infections.

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