scholarly journals A Novel Automated Approach to Enabling High-Throughput Cell Line Development, Selection, and Other Cell Culture Tasks Performed in Erlenmeyer (Shake) Flasks

2008 ◽  
Vol 13 (3) ◽  
pp. 145-151 ◽  
Author(s):  
D THOMAS ◽  
T WARD ◽  
R DRAKE ◽  
R HOLLWAY ◽  
D SAVAGE ◽  
...  
2021 ◽  
Author(s):  
Marek Widera ◽  
Alexander Wilhelm ◽  
Tuna Toptan ◽  
Johanna M. Raffel ◽  
Eric Kowarz ◽  
...  

SummaryThe severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) is the causative agent of the acute respiratory disease COVID-19, which has become a global concern due to its rapid spread. The common methods to monitor and quantitate SARS-CoV-2 infectivity in cell culture are so far time-consuming and labor-intensive. Using the Sleeping Beauty transposase system, we generated a robust and versatile reporter cell system that allows SARS-CoV-2 infection experiments compatible for high-throughput and live cell imaging. The reporter cell is based on lung derived A549 cells, which show a profound interferon response and convenient cell culture characteristics. ACE2 and TMPRSS2 were introduced for constitutive expression in A549 cells. Subclones with varying levels of ACE2/TMPRSS2 were screened for optimal SARS-CoV2 susceptibility. Furthermore, extensive evaluation demonstrated that SARS-CoV-2 infected reporter cells were distinguishable from mock-infected cells and already showed approximately 12 h post infection a clear signal to noise ratio in terms of cell roughness, fluorescence and a profound visible cytopathic effect. Moreover, due to the high transfection efficiency and proliferation capacity, Sleeping Beauty transposase-based overexpression cell lines with a second inducible fluorescence reporter cassette (eGFP) can be generated in a very short time, enabling the investigation of host and restriction factors in a doxycycline-inducible manner. Thus, the novel reporter cell line allows rapid and sensitive detection of SARS-CoV-2 infection and the screening for host factors essential for viral replication.Highlights- Sleeping Beauty transposon-based cellular system was used to generate a highly susceptible cell line for monitoring SARS-CoV-2 infection- The versatile reporter cell line A549-AT is suitable for rapid and sensitive high-throughput assays- Additional gene specific expression cassettes allow the identification of SARS-CoV-2 host dependency and restriction factors


2016 ◽  
Vol 8 (39) ◽  
pp. 7193-7198 ◽  
Author(s):  
Kanvasri N. Jonnalagadda ◽  
Lam Raga A. Markely ◽  
Bing Guan ◽  
Christina Alves ◽  
Shashi Prajapati

A high-throughput method to quantify sialic acids was developed on a ForteBio Octet QK384 platform to support cell-line development.


2020 ◽  
Author(s):  
Yong Jae Kim ◽  
Sang Kyul Han ◽  
Seongtae Yoon ◽  
Chan-Wha Kim

Abstract Recent cell culture media for mammalian cells can be abundantly formulated with nutrients supporting production, but such media can be limited to use in host cell culture, transfection, cell cloning, and cell growth under the low cell density conditions. In many cases, appropriate platform media are used for cell line development, and then replaced with rich media for production. In this study, we demonstrate rich chemically defined media for Chinese hamster ovary (CHO) cells that are suitable as basal media both for cell line development and for final production of culture process. Set up for transfection, semi-solid media optimization, mini-pool screening, and single cell cloning media development were performed, and final clones were obtained with higher productivity in fed-batch culture mode using rich formulated media comparing with lean formulated media. Developed methods may remove the requirements for cell adaptation to production media after cell line development, and relieve the clonality issues associated with changing the culture media. Furthermore, established methods have advantages over traditional approaches, including saving resources and decreasing the time and the effort required to optimize the production process.


2015 ◽  
Vol 9 (S9) ◽  
Author(s):  
Jens Niklas ◽  
Julia Retey ◽  
Steffen Fehrmann ◽  
Ludwig Macko ◽  
Thomas Hartsch ◽  
...  

2020 ◽  
Vol 92 (3) ◽  
pp. 2764-2769
Author(s):  
Fateme Tousi ◽  
Yan Jiang ◽  
Sharmila Sivendran ◽  
Yvonne Song ◽  
Susan Elliott ◽  
...  

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