Establishment of 2D Cell Cultures Derived From 3D MCF‐7 Spheroids Displaying a Doxorubicin Resistant Profile

Abstract

In vitro 3D cancer spheroids generally exhibit a drug resistance profile similar to that found in solid tumors. Due to this property, these models are an appealing for anticancer compounds screening. Nevertheless, the techniques and methods aimed for drug discovery are mostly standardized for cells cultured in 2D. The development of 2D cell culture models displaying a drug resistant profile is required to mimic the in vivo tumors, while the equipment, techniques, and methodologies established for conventional 2D cell cultures can continue to be employed in compound screening. In this work, the response of 3D-derived MCF-7 cells subsequently cultured in 2D in medium supplemented with glutathione (GSH) (antioxidant agent found in high levels in breast cancer tissues and a promoter of cancer cells resistance) to Doxorubicin (DOX) is evaluated. These cells demonstrated a resistance toward DOX closer to that displayed by 3D spheroids, which is higher than that exhibited by standard 2D cell cultures. In fact, the 50% inhibitory concentration (IC 50 ) of DOX in 3D-derived MCF-7 cell cultures supplemented with GSH is about eight-times higher than that obtained for conventional 2D cell cultures (cultured without GSH), and is only about two-times lower than that attained for 3D MCF-7 spheroids (cultured without GSH). Further investigation revealed that this improved resistance of 3D-derived MCF-7 cells may result from their increased P-glycoprotein (P-gp) activity and reduced production of intracellular reactive oxygen species (ROS).

Application of an open-chamber multi-channel microfluidic device to test chemotherapy drugs

“…Our ECM-like gel system, which could trap the cells, was expected to impose these cellular behaviors in a similar manner. This assumption was supported by the IC 50 of doxorubicin for MCF-7 cells, which was comparable to the value obtained in a recent study using a 3-D context 33 . As our on-chip drug efficacy test results adequately reflected this biology, it was possible to avoid an overestimation of potency.…”

“…This finding indicates that there was no effect in the lanes adjacent to the drug-release, even in lane 8, where the drug concentration in the adjacent lane was the highest. By comparing the calculated statistical differences (i.e., p-values) between the viable cells in lanes 3 ( www.nature.com/scientificreports/ doxorubicin was comparable to the value obtained in a recent study using a 3-D context 33 . The Live/Dead stained images in Fig.…”

Assessing the Combinatorial Chemo‐Photothermal Therapy Mediated by Sulfobetaine Methacrylate‐Functionalized Nanoparticles in 2D and 3D In Vitro Cancer Models

“…3D tumor spheroids were produced as previously described. [ 14,31 ] Briefly, agarose structures (2% (w/v)) with spherical microwells were formed by using a micro‐mold (3D Petri Dish, Microtissues Inc., Providence RI, USA). Afterward, MCF‐7 cells and NHDF at 1:1 cell ratio were seeded in the agarose structures (1 × 10 6 cells per structure), leading to the formation of 81 spheroids per agarose structure.…”

<p>Characterization of Triple-Negative Breast Cancer MDA-MB-231 Cell Spheroid Model</p>

“…For instance, the BT-474 spheroids modulate the distributions of BC stem cell population and human epidermal growth factor receptor-2 (HER2), and show a higher antiapoptotic level than those cultured in 2D monolayer. 13,14 Besides, MCF-7 spheroids reveal the role of tumor microenvironment in tumor metastasis, and they are more resistant to drug treatments than those cultured as a 2D monolayer. 15 Metastasis is a multi-step process involving numerous interactions between a tumor and its microenvironment.…”