title: Modelling the tumour microenvironment in long-term microencapsulated 3D co-cultures recapitulates phenotypic features of disease progression.
abstract: 3D cell tumour models are generated mainly in non-scalable culture systems, using bioactive scaffolds. Many of these models fail to reflect the complex tumour microenvironment and do not allow long-term monitoring of tumour progression. To overcome these limitations, we have combined alginate microencapsulation with agitation-based culture systems, to recapitulate and monitor key aspects of the tumour microenvironment and disease progression. Aggregates of MCF-7 breast cancer cells were microencapsulated in alginate, either alone or in combination with human fibroblasts, then cultured for 15 days. In co-cultures, the fibroblasts arranged themselves around the tumour aggregates creating distinct epithelial and stromal compartments. The presence of fibroblasts resulted in secretion of pro-inflammatory cytokines and deposition of collagen in the stromal compartment. Tumour cells established cell-cell contacts and polarised around small lumina in the interior of the aggregates. Over the culture period, there was a reduction in oestrogen receptor and membranous E-cadherin alongside loss of cell polarity, increased collective cell migration and enhanced angiogenic potential in co-cultures. These phenotypic alterations, typical of advanced stages of cancer, were not observed in the mono-cultures of MCF-7 cells. The proposed model system constitutes a new tool to study tumour-stroma crosstalk, disease progression and drug resistance mechanisms.
authors.affiliation: iBET, Instituto de Biologia Experimental e Tecnológica, Oeiras, Portugal; Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, Oeiras, Portugal.
authors.forename: Marta F
authors.initials: MF
authors.lastname: Estrada
authors.affiliation: iBET, Instituto de Biologia Experimental e Tecnológica, Oeiras, Portugal; Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, Oeiras, Portugal.
authors.forename: Sofia P
authors.initials: SP
authors.lastname: Rebelo
authors.affiliation: European Cancer Stem Cell Institute, Cardiff University, Cardiff, UK; Bioscience, Oncology iMed, AstraZeneca, Cheshire, UK.
authors.forename: Emma J
authors.initials: EJ
authors.lastname: Davies
authors.affiliation: Institute of Molecular Pathology and Immunology, University of Porto (IPATIMUP), Portugal.
authors.forename: Marta T
authors.initials: MT
authors.lastname: Pinto
authors.affiliation: iBET, Instituto de Biologia Experimental e Tecnológica, Oeiras, Portugal; Cell Imaging Unit, Instituto Gulbenkian de Ciência, Oeiras, Portugal.
authors.forename: Hugo
authors.initials: H
authors.lastname: Pereira
authors.affiliation: iBET, Instituto de Biologia Experimental e Tecnológica, Oeiras, Portugal; Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, Oeiras, Portugal.
authors.forename: Vítor E
authors.initials: VE
authors.lastname: Santo
authors.affiliation: European Cancer Stem Cell Institute, Cardiff University, Cardiff, UK.
authors.forename: Matthew J
authors.initials: MJ
authors.lastname: Smalley
authors.affiliation: Bioscience, Oncology iMed, AstraZeneca, Cheshire, UK.
authors.forename: Simon T
authors.initials: ST
authors.lastname: Barry
authors.affiliation: Cell Imaging Unit, Instituto Gulbenkian de Ciência, Oeiras, Portugal.
authors.forename: Emilio J
authors.initials: EJ
authors.lastname: Gualda
authors.affiliation: iBET, Instituto de Biologia Experimental e Tecnológica, Oeiras, Portugal; Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, Oeiras, Portugal.
authors.forename: Paula M
authors.initials: PM
authors.lastname: Alves
authors.affiliation: Boehringer Ingelheim RCV, Wien, Austria.
authors.forename: Elizabeth
authors.initials: E
authors.lastname: Anderson
authors.affiliation: iBET, Instituto de Biologia Experimental e Tecnológica, Oeiras, Portugal; Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, Oeiras, Portugal. Electronic address: anabrito@itqb.unl.pt.
authors.forename: Catarina
authors.initials: C
authors.lastname: Brito
mesh_terms: D018920:Coculture Techniques; D018450:Disease Progression; D019008:Drug Resistance, Neoplasm; D006801:Humans; D061986:MCF-7 Cells; D059016:Tumor Microenvironment
keywords: 3D; Alginate microencapsulation; Co-culture; Stirred-tank bioreactors; Tumour microenvironment; Tumour progression