Decellularised extracellular matrix as a platform for cancer modelling

Abstract

Event Abstract Back to Event Decellularised extracellular matrix as a platform for cancer modelling Naledi Shologu1, 2, Abhay Pandit2 and Dimitrios Zeugolis1, 2 1 National University of Ireland Galway (NUIG), Regenerative, Modular & Developmental Engineering Laboratory (REMODEL), Ireland 2 National University of Ireland Galway (NUIG), Centre for Research in Medical Devices (CÚRAM), Ireland Introduction: Existing approaches aim to mimic the in vivo microenvironment by establishing the cell–cell and cell–matrix contact through various cell culture systems namely, hydrogels, fibres, sponges, microfluidics, nano-texturing, surface-coating etc[1]. Conversely, there is an inefficiency of the cells to form their own microenvironment in vitro as a result of unsubstantial amounts of deposited extracellular matrix (ECM) within a useful time frame. To solve these problems, a biophysical approach, termed macromolecular crowding (MMC), that increases thermodynamic activities and biological processes by several orders of magnitude, is proposed as means to create ECM-based scaffold as a bases for an in vitro cancer models. The principle of MMC is derived from the notion that in vivo cells reside in a highly crowded/dense extracellular space, whilst in vitro they are cultured under very dilute conditions that prohibit normal cell function. Recent work demonstrates that under MMC conditions, scaffold-free equivalents can be produced within 2-6 days in culture, whilst in the absence of MMC tissue constructs of similar thickness would have taken several months to culture[2]. Using this phenomenon, a cell-derived ECM can be fabricated in vitro within a timely manner. After the cell-derived ECM has been facilitated, cell-synthesized matrices can be utilised as a new cell-formed substrate after a decellularization treatment[3]. Materials and Methods: Human adipose-derived stem cells (hADSC) and human adult mammary fibroblasts (hMF) were cultured for 2, 7, 10, 14 and 21 days with various carrageenan (MMC) concentrations in normoxia (21%) at 37ºC and 5% CO2 in a humidified atmosphere and after the ECM were decellularized and recellularized with breast cancer cell lines. Total collagen and protein quantification deposition was assessed using sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE), hydroxyproline assay and BCA protein assay. Cellular function was assessed namely: cell metabolic activity (alamarBlue® assay), cell viability (LIVE/DEAD® assay), and cell proliferation (PicoGreen® Assay). Gelatine Zymography was used to assess matrix metalloproteinase (MMPs). Conclusions: MMC can be used effectively to accelerate ECM deposition by hADSCs and hMFs, to facilitate production of matrix-rich 3D tumour models compared to 2D monolayer model, for the in vitro assessment of anti-cancer drug response. This dynamic approach paves the ways for the development of a biologically relevant scaffold-free assay system feasible as a robust preclinical anti-cancer drug screening platform. The authors would like to acknowledge the College of Engineering and Informatics, NUIG for financial support