High Content Imaging (HCI) on Miniaturized Three-Dimensional (3D) Cell Cultures.

Moo-Yeal Lee,Pranav Joshi

doi:10.3390/bios5040768

Abstract

High content imaging (HCI) is a multiplexed cell staining assay developed for better understanding of complex biological functions and mechanisms of drug action, and it has become an important tool for toxicity and efficacy screening of drug candidates. Conventional HCI assays have been carried out on two-dimensional (2D) cell monolayer cultures, which in turn limit predictability of drug toxicity/efficacy in vivo; thus, there has been an urgent need to perform HCI assays on three-dimensional (3D) cell cultures. Although 3D cell cultures better mimic in vivo microenvironments of human tissues and provide an in-depth understanding of the morphological and functional features of tissues, they are also limited by having relatively low throughput and thus are not amenable to high-throughput screening (HTS). One attempt of making 3D cell culture amenable for HTS is to utilize miniaturized cell culture platforms. This review aims to highlight miniaturized 3D cell culture platforms compatible with current HCI technology.

Highlights

Current high-throughput screening (HTS) technology, capable of screening a large number of new compounds against an increasing number of new targets, evaluates a single endpoint involved in drugBiosensors 2015, 5 efficacy and toxicity
Due to the limited scope of our review, we focus on 3. Macroscale Three-Dimensional (3D) cell culture technologies that can be applicable to high content imaging (HCI)
HCI assays have been demonstrated in PDMS microwells with immune cells that were monitored in real-time to evaluate cell viability, membrane potential (MMP), reactive oxygen species (ROS)

Summary

Introduction

Current high-throughput screening (HTS) technology, capable of screening a large number of new compounds against an increasing number of new targets, evaluates a single endpoint involved in drug. This approach often lacks the ability to provide highly predictive information on drug responses in vivo, which is critical to reduce the high attrition rate in downstream drug discovery pipelines. To address this issue, high content imaging (HCI) or high content screening (HCS) technology—which refers to a high-throughput, automated microscope-based assay that provides information on multiple properties or features of individual cells simultaneously with several fluorescent dyes—has been adapted to a more systematic and accurate evaluation of drug candidates [1,2]. A suite of HCI assays have been developed to assess mechanisms of compound toxicity and evaluate the effects on organ toxicities, such as hepatotoxicity, cardiotoxicity, and neurotoxicity (Table 2)

Miniaturized 3D Cell Culture Systems and Their Application in HCI

Microwells

Cellular Microarrays

Microfluidic Devices

Conclusions

Conflicts of Interest