Novel Phenotypic Fluorescent Three-Dimensional Co-Culture Platforms for Recapitulating Tumor in vivo Progression and for Personalized Therapy

Abstract

Because three-dimensional (3D) in vitro models are more accurate than 2D cell culture models and faster and cheaper than animal models, they have become a prospective trend in the biomedical and pharmaceutical fields, especially for personalized and targeted therapies. Because appropriate 3D models can be customized to mimic the in vivo microenvironment wherein various cell populations grow within an intricate but well organized extracellular matrix (ECM), they can accurately recapitulate physiological and pathophysiological progressions. The majority of cancers are carcinomas, which originate from epithelial cells, and dynamically interact with non-malignant cells including stromal cells (fibroblasts), vascular cells (endothelial cells and pericytes), immune cells (macrophages and mast cells), and the ECM. Employing a tumor monoclonal colony, tumor xenograft or patient cancer biopsy into an in vivo-like microenvironment, the native signaling pathways, cell-cell and cell-matrix interactions, and cell phenotypes are preserved and our fluorescent phenotypic 3D co-culture platforms can then accurately recapitulate the tumor in vivo scenario including tumor induced angiogenesis, tumor growth, and metastasis.In this paper, we describe a robust and standardized method to co-culture a tumor colony or biopsy with different cell populations, e.g., endothelial cells, immune cells, pericytes, etc. The procedures for recovering cells from the co-culture for molecular analyses, imaging, and analyzing are also described. We selected ECM solubilized extract derived from Engelbreth-Holm-Swam sarcoma cells. Because the 3D co-culture platforms can provide drug chemosensitivity data within 9 days that is equivalent to the results generated from mouse tumor xenograft models in 50 days, the 3D co-culture platforms are more accurate, efficient, and cost-effective and may replace animal models in the near future to predict drug efficacy, personalize therapies, prevent drug resistance, and improve the quality of life.

Differential effects of sulforaphane in regulation of angiogenesis in a co-culture model of endothelial cells and pericytes

“…Conventional planar cultures fail to recreate the in vivo physiology of the microvasculature with respect to 3D geometry (lumens and axial branching points) and interactions of endothelium with perivascular cells, extracellular tissue and blood flow (40,41). There is strong evidence that the 3D in vitro model consisting of multiple stromal cells is not only cheaper but also provides quicker results than animal models.…”

Extracellular Matrix Modulates Angiogenesis in Physiological and Pathological Conditions

“…The three-dimensional (3D) and computational in vitro studies [ 1 , 2 ] clearly demonstrate that besides its remodelling ECM controls and regulates physiological and pathological angiogenesis [ 3 ] at several levels by several ways.…”

Automated tracking of tumor-stroma morphology in microtissues identifies functional targets within the tumor microenvironment for therapeutic intervention

“…The major challenge regarding complex 3D cell cultures is the detailed analysis of the experiments, including segmentation and tracking of cell movements as well as the analysis of their distinct morphologies [ 3 , 15 ]. Most analyses of 3D cultures that include stromal components only provide poorly informative growth curves from generalized fluorescent measurements or impedance, sometimes combined with incidental, molecular snapshots by immunofluorescence (IF) end-point staining [ 16 – 21 ]. Alterations in stromal motility and tumor cell plasticity are difficult to measure and usually ignored.…”