A novel pancreatic tumour and stellate cell 3D co-culture spheroid model.

Autor: Norberg, K. J., Liu, X., Fernández Moro, C., Strell, C., Nania, S., Blümel, M., Balboni, A., Bozóky, B., Heuchel, R. L., Löhr, J. M.
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Zdroj: BMC Cancer; 5/27/2020, Vol. 20 Issue 1, p1-13, 13p, 1 Black and White Photograph, 6 Graphs
Abstrakt: Background: Pancreatic ductal adenocarcinoma is a devastating disease with poor outcome, generally characterized by an excessive stroma component. The purpose of this study was to develop a simple and reproducible in vitro 3D-assay employing the main constituents of pancreatic ductal adenocarcinoma, namely pancreatic stellate and cancer cells.Method: A spheroid assay, directly co-culturing human pancreatic stellate cells with human pancreatic tumour cells in 3D was established and characterized by electron microscopy, immunohistochemistry and real-time RT-PCR. In order to facilitate the cell type-specific crosstalk analysis by real-time RT-PCR, we developed a novel in vitro 3D co-culture model, where the participating cell types were from different species, human and mouse, respectively. Using species-specific PCR primers, we were able to investigate the crosstalk between stromal and cancer cells without previous cell separation and sorting.Results: We found clear evidence for mutual influence, such as increased proliferation and a shift towards a more mesenchymal phenotype in cancer cells and an activation of pancreatic stellate cells towards the myofibroblast phenotype. Using a heterospecies approach, which we coined virtual sorting, confirmed the findings we made initially in the human-human spheroids.Conclusions: We developed and characterized different easy to set up 3D models to investigate the crosstalk between cancer and stroma cells for pancreatic cancer. [ABSTRACT FROM AUTHOR]
Databáze: Complementary Index
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