Abstract

Performing drug screening of tissue derived from cancer patient biopsies using physiologically relevant 3D tumour models presents challenges due to the limited amount of available cell material. Here, we present a microfluidic platform that enables drug screening of cancer cell-enriched multicellular spheroids derived from tumour biopsies, allowing extensive anticancer compound screening prior to treatment. This technology was validated using cell lines and then used to screen primary human prostate cancer cells, grown in 3D as a heterogeneous culture from biopsy-derived tissue. The technology enabled the formation of repeatable drug concentration gradients across an array of spheroids without external fluid actuation, delivering simultaneously a range of drug concentrations to multiple sized spheroids, as well as replicates for each concentration. As proof-of-concept screening, spheroids were generated from two patient biopsies and a panel of standard-of-care compounds for prostate cancer were tested. Brightfield and fluorescence images were analysed to provide readouts of spheroid growth and health, as well as drug efficacy over time. Overall, this technology could prove a useful tool for personalised medicine and future drug development, with the potential to provide cost- and time-reduction in the healthcare delivery.

Highlights

  • Over the last 10 years, the use of three-dimensional (3D) cell culture models and consideration of the extracellular microenvironment have been shown to be of immense importance when studying cancer therapies and the mechanisms leading to drug resistance and metastasis[1,2,3]

  • Microfluidic technologies have increasingly been used for spheroid-based assays and offer viable solutions when working with biopsy-derived tumour tissue, providing precise control over the cellular microenvironment, 3D cell culture and medium- to high-throughput readouts in a cost-effective manner[16,17,18,19]

  • We present a microfluidic platform and associated cell culture protocols, aimed to facilitate extensive screening of cancer cell-enriched multicellular spheroids derived from human tumour biopsies

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Summary

Introduction

Over the last 10 years, the use of three-dimensional (3D) cell culture models and consideration of the extracellular microenvironment have been shown to be of immense importance when studying cancer therapies and the mechanisms leading to drug resistance and metastasis[1,2,3]. Novel aspects of our microfluidic design include self-generating perfusion of nutrients and repeatable, long-lasting drug concentration gradients across an array of hundreds of spheroids without requiring equipment to be connected to the device for fluid actuation Using this system, several concentration response curves (8-point curves obtained from a starting sample of ~50,000 cells, with each point averaging 10–24 spheroids) were obtained per biopsy, providing information on drug efficacy over time from the analysis of brightfield and fluorescence images. Several concentration response curves (8-point curves obtained from a starting sample of ~50,000 cells, with each point averaging 10–24 spheroids) were obtained per biopsy, providing information on drug efficacy over time from the analysis of brightfield and fluorescence images Overall, this is the first example of an equipment-free, lab-on-a-chip platform enabling miniaturised compound screening on 3D tumour models from biopsy tissue with a throughput that is 100-fold greater than any previously described for spheroids overall[8,23,24,25,26]. This novel approach offers an animal-free, practical solution to personalised drug screening of 3D tumour biopsy preparations

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