Oral Presentation Symposium on Proteases and the Tumouri Microenvironment 2017

A 3D patient-derived bioengineered model to investigate the stromal-epithelial-extracellular matrix interactions during prostate cancer initiation and progression (#3)

Elena M De-Juan-Pardo 1 , Maria Flandes-Iparraguirre 1 , Laura Bray 1 , Brooke A Pereira 2 , Stuart J Ellem 2 , Mitchell G Lawrence 2 , Gail P Risbridger 2 , Dietmar W Hutmacher 1 3
  1. Centre for Regenerative Medicine, Institute of Health and Biomedical Innovation (IHBI), Queensland University of Technology (QUT), Kelvin Grove, QLD, Australia
  2. Department of Anatomy and Developmental Biology, Biomedicine Discovery Institute, Monash University, Clayton, VIC, Australia
  3. ARC Centre in Additive Biomanufacturing, Queensland University of Technology (QUT), Kelvin Grove, QLD, Australia

Prostate cancer (PCa) remains one of the most frequent malignancies in men, representing an estimated 25% of all the cancer cases diagnosed. Nowadays it is widely accepted that the tumour microenvironment plays a key role in the mechanisms of PCa initiation and progression, however, there is a lack of models that successfully incorporate the multiple components of the tumour microenvironment.
In this work, we developed a three-dimensional (3D) bioengineered in vitro model of early stage PCa using patient-derived stromal cells, 3D printed scaffolds and hydrogels. This platform incorporates the stromal compartment, the epithelial compartment, and the basement membrane in between. In order to achieve this, primary human normal or cancer-associated fibroblasts (NPF or CAF, respectively) were first grown until confluency on a melt electrospun medical grade poly(ε-caprolactone) scaffold, and allowed to form their own dense extracellular matrix (ECM). In parallel, the epithelial compartment was prepared by growing benign prostatic hyperplasia epithelial cells (BPH-1) until confluency onto a thin gelatin methacryloyl hydrogel reinforced with a highly porous melt electrospun mesh. Finally, the epithelial compartment and the confluent scaffold were co-cultured together until a rich basement membrane was formed between the stromal and epithelium compartment.
The data confirmed the deposition of a distinct 3D ECM by CAF or NPF within the scaffolds. Results showed that BPH-1-NPF co-cultures, but not BPH-1-CAF co-cultures, formed a highly organized basement membrane, as confirmed by positive immunofluorescent and immunohistochemical staining of laminin and collagen IV. Transmission electron microscopy was also used to further investigate the morphological features of the basement membrane.
In summary, we have established a robust, physiologically relevant 3D in vitro model of early stage PCa that incorporates the basement membrane and patient-derived stromal cells to dissect the stromal-epithelial-ECM interactions in the initiation and progression of this widespread male malignancy.