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- Title
A novel pancreatic tumour and stellate cell 3D co-culture spheroid model.
- Authors
Norberg, K. J.; Liu, X.; Fernández Moro, C.; Strell, C.; Nania, S.; Blümel, M.; Balboni, A.; Bozóky, B.; Heuchel, R. L.; Löhr, J. M.
- Abstract
<bold>Background: </bold>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.<bold>Method: </bold>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.<bold>Results: </bold>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.<bold>Conclusions: </bold>We developed and characterized different easy to set up 3D models to investigate the crosstalk between cancer and stroma cells for pancreatic cancer.
- Subjects
CANCER cells; CELL separation; ELECTRONIC money; PANCREATIC cancer; TUMORS; PANCREATIC tumors; RESEARCH methodology; IMMUNOHISTOCHEMISTRY; TISSUE culture; CELL physiology; DUCTAL carcinoma; ELECTRON microscopy; CELL communication; CELLS; CELL lines; POLYMERASE chain reaction; PHENOTYPES
- Publication
BMC Cancer, 2020, Vol 20, Issue 1, p1
- ISSN
1471-2407
- Publication type
journal article
- DOI
10.1186/s12885-020-06867-5