Abstract
The development of three-dimensional (3D) cultures is increasing, as they are able to provide the utility of in vitro models and the strength of testing in physiologically relevant systems. When cultured in a scaffold-free agarose hydrogel system, MCF-7 human breast carcinoma cells organize and develop into microtissues that contain a luminal space, in stark contrast to the flat morphology of MCF-7 two-dimensional (2D) monolayer cultures. Following exposure to 1nM E2, expression of typical estrogen-responsive genes, including progesterone receptor (PGR), PDZ containing domain 1 (PDZK1) and amphiregulin (AREG) is increased in both 2D and 3D cultures. When examining expression of other genes, particularly those involved in cell adhesion, there were large changes in 3D MCF-7 microtissues, with little to no change observed in the MCF-7 monolayer cultures. Together, these results indicate that while the initial estrogen-regulated transcriptional targets respond similarly in 2D and 3D cultures, there are large differences in activation of other pathways related to cell-cell interactions.
Highlights
As a result of the National Research Council report “Toxicity Testing in the 21st Century: A Vision and a Strategy,” [1] there is a large need to develop more efficient and physiologically relevant models for evaluation of compound safety and toxicity
While 2D cultures are technically easy to conduct and inexpensive, they do not recapitulate the biology of cells in vivo, where tissues are highly cell dense and cells are in contact with other cell types and extracellular matrix (ECM)
Complete gene expression results are reported in S2 Table
Summary
As a result of the National Research Council report “Toxicity Testing in the 21st Century: A Vision and a Strategy,” [1] there is a large need to develop more efficient and physiologically relevant models for evaluation of compound safety and toxicity. Due to the cost- and timeintensive nature of animal testing, in vitro based screening models are growing in favor. In vitro cell culture models allow for the use of human cell lines, enabling researchers to better evaluate human health and disease states when compared to in vivo animal models. While 2D cultures are technically easy to conduct and inexpensive, they do not recapitulate the biology of cells in vivo, where tissues are highly cell dense and cells are in contact with other cell types and extracellular matrix (ECM). Previous work has demonstrated that this contact with ECM and cells can alter gene expression profiles and PLOS ONE | DOI:10.1371/journal.pone.0157997 July 5, 2016
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