Overview of 3D Cell Culture Model Systems and Factors to Consider When Choosing and Validating Cell-Based Assays for Use With 3D Cultures
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Dr. Terry Riss started the Cell Biology program at Promega Corporation in 1990 and held several R&D and Project Management positions since. Dr. Riss managed development of cell viability, cytotoxicity, apoptosis, and protease assay systems and also lead efforts to identify and promote multiplexing of cell-based assays to determine the mechanism of cell death. Dr. Riss now serves as Senior Product Specialist, Cell Health involved in outreach educational training activities including validating assay systems applied to 3D cell culture models.
Terry Riss, Ph.D.
Senior Product Specialist, Cell Health
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- Original Webinar Date: Tuesday, March 11, 2014
This webinar will describe an overview of various 3D cell culture model systems including advantages and disadvantages of each model. Also described will be examples of cell-based assays originally designed for monolayer cultures with optimized protocols for use with 3D cultures. View full abstract below.
- View full abstract below.
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 the 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 also be presented.