Factors to Consider When Choosing Cell-Based Assays for Use with 3D Cultures
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United States of America
16 December 2014
|Description:||This webinar is sponsored by Corning Life Sciences and presented by Promega Corporation describing the advantages and disadvantages of various 3D culture methods|
|References:||ppt slides: https://promega.media/-/media/files/promega-worldwide/worldwide/factors-to-consider-when-choosing-cell-based-assays-for-use-with-3d-cultures.pdf?la=en|
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In vitro methods
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Cells cultured in 3D model systems often acquire relatively large in vivo-like structures compared to the thickness of a 2D monolayer of cells grown on standard plastic plates. Multicellular 3D culture systems containing more than one cell type and exhibiting formation of a complex extracellular matrix represent a more physiologically relevant environment, yet provide a challenge for assay chemistries originally designed for measuring events from monolayers of cells. There is an unmet need for guidelines for design and verification of convenient and effective assays useful for larger 3D microtissues. Critical factors to consider for each model system and cell type include effective penetration of detection reagents and/or complete lysis of microtissue structures using combinations of detergent and physical disruption. We will present the approach used to verify performance of a bioluminescent ATP detection assay for measuring cell viability, a caspase assay for detecting apoptosis, and cell stress reporter assays to detect mechanisms leading to cytotoxicity. Recommendations for factors to consider when verifying performance of cell health assays on 3D culture models will be presented.
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