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.

Identification of an easy to use 3D culture model to investigate invasion and anticancer drug response in chondrosarcomas

“…For instance, Brito and collaborators developed alginate embedded culture of epithelial tumor cells aggregated together with human fibroblasts [33]. Such modifications and co-cultures may be made, in the future, to improve this 3D culture system in order to get again closer to natural tumoral microenvironment.…”

PEG‐fibrinogen hydrogels for three‐dimensional breast cancer cell culture

“…In recent studies, 3D breast cancer models have been developed using various biomimetic materials for the investigation of tumorigenic phenomena such as cellular migration, angiogenesis, and metastasis and for anti‐cancer drug testing . Generally, breast cancer can be classified into three distinct molecular subtypes: luminal subtype (estrogen receptor positive (ER+)/progesterone receptor positive (PR+)/human epidermal growth factor receptor negative (HER2‐)), HER‐2 subtype (ER‐/PR‐/HER2+), and triple negative subtype (ER‐/PR‐/HER2‐).…”

“…The development of three-dimensional (3D) tissueengineered cancer models employing the use of biomimetic polymers for cell encapsulation and culture is an important step toward in vitro modeling of the disease for investigations of tumorigenic mechanisms and drug-testing applications. [1][2][3][4] In traditional drug testing approaches, cancer cells cultured on two-dimensional (2D) substrates are used for testing efficacies of potential candidate drugs. However, due to inherent differences between 2D cultured cells and those in native tumor tissue, many compounds fail to meet desired levels of efficacy and safety in clinical studies.…”

“…In recent studies, 3D breast cancer models have been developed using various biomimetic materials for the investigation of tumorigenic phenomena such as cellular migration, angiogenesis, and metastasis and for anti-cancer drug testing. 1,2,[25][26][27][28][29][30] Generally, breast cancer can be classified into three distinct molecular subtypes: luminal subtype (estrogen receptor positive (ER1)/progesterone receptor positive (PR1)/human epidermal growth factor receptor negative (HER2-)), HER-2 subtype (ER-/PR-/HER21), and triple negative subtype (ER-/PR-/HER2-). In some previous studies, comparative 3D in vitro models representative of these subtypes have been developed; 2,25,[31][32][33][34] however, the synergistic role of matrix stiffness and physical properties in modulating the morphology and behavior of these multiple cellular subtypes needs to be investigated in more detail.…”

Cell spheroids: the new frontiers in in vitro models for cancer drug validation

“…Although remarkable progress has been done in in vitro anticancer therapeutics screening using 3D matrices [4], organoids [5],…”