Standardization and optimization of cell-plating densities

Thursday, October 27, 2011 — Core Poster Session
10:00 a.m. – Noon	South Lobby of Building 10	NIAID	CORE-7

Authors

• N Deiuliis
• K Lamberton
• D Opishinski
• J Michelotti

Abstract

The Integrated Research Facility (IRF) cell culture laboratory centralizes tissue culture and cell-based assay development for translational virology research. One task of the laboratory is the delivery of plated mammalian cells to standard biosafety level-2 and high containment (BSL-3 and BSL-4) laboratories. When these cells are used for plaque assays or virus preparations, it is important to control the density of the monolayers. Although confluence of monolayers is an objective measure, the interpretation is subjective. The goal of this study is to create a standard measure of cell confluence to be used in collaborative environments. We created a visual database of cell seeding number versus confluence of adherent cell lines at the IRF. By using an automated cell counter we were able to eliminate human error and ensure consistency and accuracy. Cell lines were plated at different densities, then imaged using the Incucyte™ to objectively measure confluence and determine cell-specific growth curves. Figures show cell lines plated in T175 flasks and 6-well plates at multiple seeding numbers after 1 and 2 day incubation periods. The resulting database will give virologists visual guidelines to use when ordering cells so that their experiments will be consistent and generate quality data.

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