The Microwell-mesh: A high-throughput 3D prostate cancer spheroid and drug-testing platform

Abstract

Treatment following early diagnosis of Prostate cancer (PCa) is increasingly successful, whilst the treatment of advanced and metastatic PCa remains challenging. A major limitation in the development of new therapies is the prediction of drug efficacy using in vitro models. Classic in vitro 2-dimensional (2D) cell monolayer cultures are hypersensitive to anti-cancer drugs. As a result, there has been a surge in the development of platforms that enable three dimensional (3D) cultures thought to better replicate natural physiology and better predict drug efficacy. A deficiency associated with most 3D culture systems is that their complexity reduces the number of replicates and combination therapies that can be feasibly evaluated. Herein, we describe the use of a microwell platform that utilises a nylon mesh to retain 3D micro-tumours in discrete microwells; termed the Microwell-mesh. The Microwell-mesh enables the manufacture of ~150 micro-tumours per well in a 48-well plate, and response to anti-tumour drugs can be readily quantified. Our results demonstrate that 3D micro-tumours, unlike 2D monolayers, are not hypersensitive to Docetaxel or Abiraterone Acetate, providing a superior platform for the evaluation of sequential drug treatment. In summary, the Microwell-mesh provides an efficient 3D micro-tumour platform for single and sequential drug screening.

Using high throughput microtissue culture to study the difference in prostate cancer cell behavior and drug response in 2D and 3D co-cultures

“…For example, hydrogel matrix-based 3D cultures can be costly, they commonly suffer from significant 3D tissue size heterogeneity, and harvest from the gel is necessary for many forms of analysis [ 14 ]. Our team previously introduced the Microwell-mesh as a high throughput platform suitable for 3D tissue culture [ 15 , 16 ]. The Microwell-mesh uses a microwell platform to facilitate the manufacture of hundreds of uniform multicellular 3D microtissues.…”

3D arrays of microcages by two-photon lithography for spatial organization of living cells

“…2D cell cultures with microwells entrapping air bubbles for patterning the microenvironment were demonstrated by Goral and coworkers [18]. Microwell-meshes for mimicking 3D environment were tested for anti-drug screening [19].…”

A Microfluidic Spheroid Culture Device with a Concentration Gradient Generator for High-Throughput Screening of Drug Efficacy

“…These methods rely on the self-aggregation of cells. Moreover, these 3D cell arrays would include scaffolds, such as hydrogels and meshes, to mimic cell-to-extracellular matrix interactions and tissue-specific properties [14,15]. These micro-plate-based 3D cell arrays involve robotics, liquid handling devices, sensitive detectors, and software, for data processing and control [16].…”