Shannon M. Hawkins, Bishnubrata Patra, Muhammad Abdul Lateef, Melica Nourmoussavi Brodeur, Hubert Fleury, Euridice Carmona, Benjamin Péant, Diane Provencher, Anne-Marie Mes-Masson, Thomas Gervais
Article (2020)
Abstract
Epithelial ovarian cancer (EOC) is the most lethal gynecologic malignancy in North America, underscoring the need for the development of new therapeutic strategies for the management of this disease. Although many drugs are pre-clinically tested every year, only a few are selected to be evaluated in clinical trials, and only a small number of these are successfully incorporated into standard care. Inaccuracies with the initial in vitro drug testing may be responsible for some of these failures. Drug testing is often performed using 2D monolayer cultures or 3D spheroid models. Here, we investigate the impact that these different in vitro models have on the carboplatin response of four EOC cell lines, and in particular how different 3D models (polydimethylsiloxane-based microfluidic chips and ultra low attachment plates) influence drug sensitivity within the same cell line. Our results show that carboplatin responses were observed in both the 3D spheroid models tested using apoptosis/cell death markers by flow cytometry. Contrary to previously reported observations, these were not associated with a significant decrease in spheroid size. For the majority of the EOC cell lines (3 out of 4) a similar carboplatin response was observed when comparing both spheroid methods. Interestingly, two cell lines classified as resistant to carboplatin in 2D cultures became sensitive in the 3D models, and one sensitive cell line in 2D culture showed resistance in 3D spheroids. Our results highlight the challenges of choosing the appropriate pre-clinical models for drug testing.
S1 Fig. 7-AAD and Annexin-V flow cytometry analysis. Graphs represent an example of flow cytometry analysis of spheroids treated with 300 μM carboplatin and its respective control. After excluding cell debris, viable cells were selected based on the absence of 7-AAD and/or Annexin-V markers (red square). Apoptotic and/or dead cells are shown in the other quadrants. https://doi.org/10.1371/journal.pone.0244549.s001 (TIF);
S2 Fig. Spheroid formation using hanging drops with different EOC cell lines. (A) Bright field images at day 4. (B) Bright field images at day 6. Scale bar, 500 μm. https://doi.org/10.1371/journal.pone.0244549.s002 (TIF);
S3 Fig. Spheroid formation using ULA round-bottom wells (without Matrigel) with different EOC cell lines. Bright field images at day 2. Note that TOV112D cells in these conditions form multiple spheroids of smaller size. Scale bar, 300 μm. https://doi.org/10.1371/journal.pone.0244549.s003 (TIF)
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