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Factors to Consider for Adapting Off-the-Shelf Assays for use with 3D Culture Models

Part # PS327

Abstract

Terry Riss, Sarah Duellman, Kevin Kupcho, Brad Hook, Samantha Lewis, Mike Valley, Chad Zimprich, Andrew Niles and Dan Lazar
Promega Corporation, 2800 Woods Hollow Rd, Madison, WI, 53711

Most commercially available assay kits to measure cell viability, cytotoxicity, apoptosis, reporter gene activity, etc. were designed using monolayer cultures that may be only 5μm thick. However, 3D cell culture models have much larger physical dimensions that may require 100s of μm of reagent penetration.There is an unmet need for assays that have been validated to measure markers in larger 3D culture models. We present examples of changing the reagent formulation and/or the assay protocol to improve the ability to lyse cells. We also describe adapting the use of cell permeable and nonpermeable small molecule probes to detect viable and dead cells in real time assays.

Printed in USA.