Three-Dimensional Cell Cultures in Drug Discovery and Development

Abstract

The past decades have witnessed significant efforts toward the development of three-dimensional (3D) cell cultures as systems that better mimic in vivo physiology. Today, 3D cell cultures are emerging, not only as a new tool in early drug discovery but also as potential therapeutics to treat disease. In this review, we assess leading 3D cell culture technologies and their impact on drug discovery, including spheroids, organoids, scaffolds, hydrogels, organs-on-chips, and 3D bioprinting. We also discuss the implementation of these technologies in compound identification, screening, and development, ranging from disease modeling to assessment of efficacy and safety profiles.

Keywords: 3D bioprinting; 3D cell culture; disease models; efficacy; multicellular spheroid; organoids; organs-on-chips; safety; screening; toxicity.

Figure 1.

Four different techniques used for spheroid cultures. (a) A well of low-adhesion plates that have a round bottom with an ultralow cell attachment coating. (b) A droplet of hanging drop plate where cells are partitioned and self-organized into a spheroid. (c) Suspension culture in bioreactor where cells become self-aggregated into spheroids. (d) A representative pillar of micropatterned plates where the cells are enriched on the top of the pillar to form a spheroid.

Figure 2.

How different three-dimensional culture techniques have been implemented in different stages of drug discovery and development processes. Representative references for each application are cited in parentheses in the graph.

Figure 3.

A label-free, single-cell, real-time assay to measure the invasion of cells in a single spheroid through a three-dimensional extracellular matrix (Matrigel). (a) Principle of the assay, which consists of four critical steps: coating the biosensor surface with Matrigel; adding medium to the well; transferring a spheroid from an ultralow attachment, round-bottomed microplate and placing it onto the top Matrigel surface; and monitoring the invasion of cells through the matrix and adhesion on the sensor surface in real time. (b–d) The time series dynamic mass redistribution (DMR) images before and after a single spheroid was placed onto the biosensor surface coated with 10 µL 0.1 mg/mL Matrigel: 0 min (b), 1 h (c), and 24 h (d). Spatial scale bar: 500 µm. Intensity scale bar: –500 pm to 2000 pm. (e) A DMR image taken 24 h after a spheroid was placed on the top Matrigel surface. Scale bar: 500 µm. (f) Representative pixelated real-time DMR signals for the black line indicated in (e). (g) The adhesion events versus cell types. (h) The adhesion time to reach 200 pm under different conditions. For (e–h), coating was 0.2 mg/mL Matrigel. Data represent mean ± SD for g (n = 3). ***p < 0.001. This figure is adapted from ref. 89 with permission.