THE PERFECT TECHNOLOGY TO STUDY TUMOR-STROMA INTERACTIONS
Our focus on the tumor-stroma interactions
3DProSeed ® is the ideal hydrogel platform to study the interactions between tumor cells and the surrounding tumor microenvironment.
We promote a unique approach to establish advanced co-cultures in fully synthetic hydrogels by sequentially seeding stromal cells and tumor cells. In a first step we culture various stromal cells in the synthetic hydrogel. These cells proliferate and deposit their endogenous extracellular matrix proteins and secrete soluble factors. In a second step we seed tumor cells in this preformed stromal environment and study the tumor behavior and development. This process is carried out by simply adding cell suspensions in a microtiter plate to offer maximal workflow integration.
STEP 1 Establish a synthetic tumor stromal microenvironment
The fate of malignant cancer cells is influenced by a multitude of cells (fibroblasts and other mesenchymal supporting cells, endothelial and immune cells), soluble factors and matrix proteins surrounding them. The ability to build a synthetic model of the stromal microenvironment in a screening compatible manner enables new generation cell-based assays in oncology research and discovery.
Fibroblasts and mesenchymal supporting cells form a 3D fibroblastic matrix in the hydrogel. The source of these fibroblast-like cells (tumor tissue vs healthy tissue) can affect tumor development in vitro. These stromal cells secrete extra-cellular matrix proteins which can be analyzed using microscopy or proteomics techniques thanks to the fully synthetic nature of the 3DProSeed hydrogel and can play a role in mechanisms of action of oncological therapies. Mesenchymal stem cells can be differentiated in the hydrogel to create a fat-rich environment that can influence the resistance of tumor cells against certain anti-cancer therapies.
Patient-derived cancer associated fibroblasts (CAFs) can be seeded on the 3DProSeed hydrogel plate to establish an artificial human model of the tumor stroma. Over time the CAFs develop a dense three-dimensional fibroblastic network that can be used to study interactions between adenocarcinoma cells and CAF, without the interference of exogenous factors and proteins.
Another important constituent of the tumor microenvironment are endothelial cells (vascular or lymphatic), which co-cultured with mesenchymal cells form an an artificial model of the vascular niche to study angiogenesis, metastasis and tumor cells' quiescence.
STEP 2 Add cancer cells to study the interaction with the stromal microenvirnoment
Tumor cells of different sources can be added to the preformed stromal microenvironment. We have experience with adenocarcinoma cell lines, primary and patient-derived cells. The video on the left shows an heterospheroid of fibroblasts and a pancreatic adenocarcinoma line seeded on a 3DProSeed gel pre-seeded with fibroblast to study invasion. Below, CAFs isolated from lung adenocarcinoma were co-cultured with patient-derived lung adenocarcinoma cells to study tumor growth and fibroblast association.
It is possible to study growth and killing of tumor cells and pinpoint the role of endogenously secreted ECM components in the tumor regulation.
Applicable read-outs are microscopy (transmission or fluorescence) immunobinding assays and proteomics.