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ConsiderationsWhen Using Frozen Cells for High Throughput Cell-Based Assays
Considerations When Using Frozen Cells for High Throughput Cell-Based Assays
Introduction and PurposeThe use of cell-based assays in High Throughput Screens
(HTS) is growing in acceptance and popularity. Advantages
of cell-based assays include: (1) distinguishing between agonist
and antagonist, (2) identification of allosteric modulators,
(3) characterization of compound attributes, such as permeability
and stability inside of cells, and, (4) assessment of
cytotoxicity.1 Ideally, the goal is to generate data that are
more physiologically predictive of the clinical outcome.
The rise in the use of HTS is predominantly due to wider
choices in technology and the availability of large batches
of cryopreserved cells. Using cryopreserved cells directly
in HTS assays allows for:
2- Improved assay performance and uniformity due to the
availability of large batches of cells at the same cell line
passage.
- Increased flexibility for running screens due to readily
available stocks of frozen cells.
- Reduced costs for screens, due to the decreased time
and reagent requirements for maintaining parallel cell
lines in culture.
Although beneficial, the use of cells (and cryopreserved
cells) for in vitro assays should be approached with caution,
as cells can demonstrate dramatically differing growth and
functional characteristics depending on environmental and
experimental conditions.
The following study investigated the impact on performance
of batch-frozen vs. continuously cultured cells in multiple
assay formats. HEK-293 and CHO-K1-M1WT2 adherent
cell lines were used in a luciferase reporter gene and calcium
flux assay, respectively. C166-GFP cells were used to assess
the impact on cellular retention on the microplate surfaces
after the various treatments. For all assay formats, the impact
of cell treatment on assay performance was assessed. The
study also examined the impact of surface chemistry on assay
performance using cryopreserved cells, comparing the standard
tissue culture surface (TCT), Corning® CellBIND®
surface and poly-D-lysine coating..
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