The Use of 3-D Cultures for High-Throughput Screening: The Multicellular Spheroid Model

Abstract

Over the past few years, establishment and adaptation of cell-based assays for drug development and testing has become an important topic in high-throughput screening (HTS). Most new assays are designed to rapidly detect specific cellular effects reflecting action at various targets. However, although more complex than cell-free biochemical test systems, HTS assays using monolayer or suspension cultures still reflect a highly artificial cellular environment and may thus have limited predictive value for the clinical efficacy of a compound. Today's strategies for drug discovery and development, be they hypothesis free or mechanism based, require facile, HTS-amenable test systems that mimic the human tissue environment with increasing accuracy in order to optimize preclinical and preanimal selection of the most active molecules from a large pool of potential effectors, for example, against solid tumors. Indeed, it is recognized that 3-dimensional cell culture systems better reflect the in vivo behavior of most cell types. However, these 3-D test systems have not yet been incorporated into mainstream drug development operations. This article addresses the relevance and potential of 3-D in vitro systems for drug development, with a focus on screening for novel antitumor drugs. Examples of 3-D cell models used in cancer research are given, and the advantages and limitations of these systems of intermediate complexity are discussed in comparison with both 2-D culture and in vivo models. The most commonly used 3-D cell culture systems, multicellular spheroids, are emphasized due to their advantages and potential for rapid development as HTS systems. Thus, multicellular tumor spheroids are an ideal basis for the next step in creating HTS assays, which are predictive of in vivo antitumor efficacy. (Journal of Biomolecular Screening 2004:273-285)

Cell aggregation enhances bone formation by human mesenchymal stromal cells

“…order to test the potential in vivo effects of drug therapies (Kobayashi et al, 1993;Kunz-Schughart, 2004), but this study establishes its potential for bone tissue engineering. Another significant advantage of the approach described in this study is that the in vitro culture time of the constructs had no statistically significant effect on the amount of bone formation after six weeks of implantation.…”

Multi‐dimensional fluorescence microscopy of living cells

“…A further prerequisite is that microscopes must be adapted to biological samples of different geometry, e.g. tissue samples or 3-dimensional cell cultures, which are more similar to tissue morphology and function than conventional 2-dimensional cell cultures on microscope slides [10].…”

Evaluation of Three-Dimensional Porous Iron-Cross-Linked Alginate as a Scaffold for Cell Culture

“…Thus, two-dimensional culture is considered to be an unnatural condition for many cell types. In order to overcome the shortcomings of two-dimensional culture, three-dimensional culture systems, such as multicellular spheroids, cellular multilayers, and matrix-embedded culture, have been devised [6]. Therefore, studies focusing on the optimal three-dimensional culture environment for cells have important implications.…”