Abstract
Purpose of this study was the development of a 3D material to be used as substrate for breast cancer cell culture. We developed composite gels constituted by different concentrations of Alginate (A) and Matrigel (M) to obtain a structurally stable-in-time and biologically active substrate. Human aggressive breast cancer cells (i.e. MDA-MB-231) were cultured within the gels. Known the link between cell morphology and malignancy, cells were morphologically characterized and their invasiveness correlated through an innovative bioreactor-based invasion assay. A particular type of gel (i.e. 50% Alginate, 50% Matrigel) emerged thanks to a series of significant results: 1. cells exhibited peculiar cytoskeleton shapes and nuclear fragmentation characteristic of their malignancy; 2. cells expressed the formation of the so-called invadopodia, actin-based protrusion of the plasma membrane through which cells anchor to the extracellular matrix; 3. cells were able to migrate through the gels and attach to an engineered membrane mimicking the vascular walls hosted within bioreactor, providing a completely new 3D in vitro model of the very precursor steps of metastasis.
Highlights
Breast cancer is the most common cancer in women across most ethnic groups and one of the leading causes of cancer-related deaths worldwide[1,2,3]
Cancer biology research has involved in vitro analysis of cell behaviour predominately using two-dimensional (2D) cell cultures and in vivo animal models[17,18]: in detail, 2D models are routinely used as initial systems for evaluating the effectiveness of molecules as potential therapeutic drugs; this initial screening precedes animal studies before advancing to human clinical trials[19]
Models and human malignancies are known: the dissimilarities in cell behaviour between 2D cultures and real tumours derive from changes in gene expression originated from the different interactions to which cells are subjected within a 2D microenvironment if compared to a more natural 3D23,24
Summary
Breast cancer is the most common cancer in women across most ethnic groups and one of the leading causes of cancer-related deaths worldwide[1,2,3]. This alginate-based model resulted appropriate for the culture of lowly aggressive cells, that both in 2D and in 3D maintain a pretty round morphology and a cluster-like organization[39], while a much more permissive environment becomes necessary when invasive phenomena need to be studied.
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