3D Cell Culture System to Study Tumour-Stroma Interactions
News May 16, 2014
The article describes a novel 3D co-culture model using non-small cell lung cancer (NSCLC) cell lines in combination with lung fibroblasts.
Automation-compatible multi-well hanging drop microtiter plates were used for the production of 3D mono- and co-cultures. In these hanging drops the two NSCLC cell lines A549 and Colo699 were cultivated either alone or co-cultured with lung fibroblasts. The viability of tumour spheroids was confirmed after five and ten days by using Annexin V/Propidium Iodide staining for flow-cytometry. Tumour fibroblast spheroid formation was characterized by scanning electron microscope (SEM), semi-thin sections, fluorescence microscope and immunohistochemistry (IHC). In addition to conventional histology, protein expression of E-Cadherin, vimentin, Ki67, fibronectin, cytokeratin 7 and α-smooth muscle actin (α-SMA) was investigated by IHC.
Lower viability was observed in A549 monocultures compared to co-cultures, whereas Colo699 monocultures showed better viability compared to co-cultures. Ki67 expression varied significantly between mono- and co-cultures in both tumour cell lines. An increase of vimentin and decreased E-Cadherin expression could be detected during the course of the cultivation suggesting a transition to a more mesenchymal phenotype. Furthermore, the fibroblast cell line showed an expression of α-SMA only in co-culture with the cancer cell line A549, thereby indicating a mesenchymal to mesenchymal shift to an even more myofibroblast phenotype.
We demonstrate that our method is a promising tool for the generation of tumour spheroid co-cultures. Furthermore, these spheroids allow the investigation of tumour-stroma interactions and a better reflection of in vivo conditions of cancer cells in their microenvironment. Our method holds potential to contribute to the development of anti-cancer agents and support the search for biomarkers.
The article, Development of an innovative 3D cell culture system to study tumour--stroma interactions in non-small cell lung cancer cells, is published in PLOS ONE and is free to access.
Using Milk Protein to 3D-Imprint Muscle and Bone CellsNews
Researchers from the University of Canterbury are replicating a 3D imprint of cells onto films made of milk protein. The films then gradually degrade, leaving the grown tissue behind.READ MORE
Cell Recycling System Offers Therapeutic Entry Point for Rare Disease TreatmentNews
Scientists have demonstrated how an investigational drug works against a rare, fatal genetic disease, Niemann-Pick type C1 (NPC1).READ MORE
Comments | 0 ADD COMMENT
EMBL Course: Transgenic Animals - Micromanipulation Techniques
Apr 10 - Apr 11, 2018
EMBO Practical Course: Extracellular Vesicles: From Biology to Biomedical Applications
Apr 09 - Apr 13, 2018
EMBO | EMBL Symposium: Tissue Self-Organisation: Challenging the Systems
Mar 11 - Mar 14, 2018
EMBL Course: Brillouin Microscopy: Emerging Tool for Probing Mechanical Properties of Living Cells
Jan 17 - Jan 19, 2018