Adapting Cell-Based Assays to the High-Throughput Screening Platform: Problems Encountered and Lessons Learned

Abstract

In recent years, cell-based phenotypic assays have emerged as an effective and robust addition to the array of assay technologies available for drug discovery in the high throughput screening arena. Previously, biochemical target-based assays have been the technology of choice. With the emergence of stem cells as a basis for a new screening technology, it is important to keep in mind the lessons that have been learned from the adaptation of existing stable cell lines onto the high throughput screening drug discovery platform, with special consideration being given to assay miniaturization, liquid handling complications and instrument-introduced artifacts. We present an overview of the problems encountered with the implementation of multiple cell-based assays at the High Throughput Screening Center at Southern Research Institute as well as empirically defined effective solutions to these problems. These include examples of artifacts induced by temperature differences throughout the screening campaign, cell plating conditions including the effect of room temperature incubation on assay consistency, DMSO carry-over, and incubator induced artifacts.

Automation of Three-Dimensional Cell Culture in Arrayed Microfluidic Devices

“…These results have created a need for higher throughput systems capable of screening the physical, chemical and biological properties of the 3D cellular microenvironment. However, adapting cell-based assays to high-throughput screening applications present challenges such as cell plating conditions, incubation times, compound addition parameters, and assay endpoint considerations that affect the quality of the final output 12 . As a result, microfabricated arrayed systems (e.g.…”

Pluripotent Human Stem Cells

“…[103] These include devising strategies for single-cell adaptation of stem cells for automated handling and high-throughput plating and screening, as well as managing the 'edge effect' of cultured cells, evaporation and gas exchange.…”

Perfusion Based Cell Culture Chips

“…Such tests have successfully been performed in batch cultures and can be scaled up to large combinatory experiments using microtiter plates and robotics [30]. Several attempts have been made to implement high-throughput cell analysis in terms of perfusion based microfluidic cell culture chips.…”