Abstract LB-111: Mastospheres as a new 3D ex vivo breast cancer microtumors for preclinical drug testing
Authors: Richon Sophie , Cottu Paul , Weiswald Louis-Bastien , Guinebretiere Jean-Marc , Marangoni Elisabetta , Assayag Franck , Thuleau Aurelie , Labiod Dalila , Decaudin Didier , Bellet Dominique , Dangles-Marie Virginie 
Published: August 2012

Abstract
There is increasing evidence that three-dimensional (3D) tissue culture technologies have many uses within the preclinical assays in cancer. The 3D tumor model accurately reproduces the in vivo tumor phenotype and represents an additional tool for studying tumor biology and allowing better preclinical evaluation of anticancer drugs. One of the used models involves small tumor aggregates, termed spheres, that are obtained from mechanical tumor dissociation and that have shown the superiority of 3D culture over standard two-dimensional cell culture for mimicking the tumor biology and drug response observed in vivo. The aim of this study was to generate new ex vivo 3D models from breast (BC) cancer xenografts established from patients’ tumor fragments. These ex vivo mastospheres are easily obtained from mechanically dissociated fresh human BC tissue xenografted in Nude mice, in a similar way as colospheres from human colon cancer (Weiswald et al, Br J Cancer 2009, 101:473). In contrast to mammospheres described in the literature, 3D mastospheres are obtained from tumor fragments, without enzymatic tumor dissociation, without matrix substratum and in SVF supplemented medium. From a large panel of patient-derived BC xenografts already well characterized (Marangoni et al, Clin Cancer Res 2007, 13:3989; Cottu et al, Breast Cancer Res Treat 2011; Reyal et al, Breast Cancer Res 2012, 14:R11), we get mastospheres from 26 out of 36 (72%) xenografts. Mastosphere formation is scored on day 1 after culture according to the number of spheres/mg of dissociated xenograft tissue. Within mastospheres, we clearly distinguish 3 distinct morphologies: round, grape-like and aggregates. Histological analyses show also that mastospheres were formed only with proliferating cancer cells. In a reproducible way, dissociation of a given xenograft leads to a similar score and to a similar morphology. All BC subtypes (luminal, triple negative and HER2+ tumors) give rise to mastospheres. We noted that the 5 out of 5 xenografts (100%) able to metastasise in Nude mice (lung metastasis) form mastospheres, suggesting that the capacity to give mastospheres could be related to tumour aggressiveness, as already reported with colospheres and colon cancer. These different features prompted us to test the potential of these mastospheres in chemosensitivity assays. We first demonstrated that mastospheres can be kept viable a couple of days, consistent with ex vivo assay aim. For this purpose, we tested the sensitivity to cisplatin in two xenograft models with different in vivo response. We found that the paired mastospheres tested in viability assays mimicked these different response profiles (mean of IC50: 1.2 µM versus 15.5 µM). In conclusion, according to these preliminary data, mastospheres deserve additional investigation because of their interest as new ex vivo microtumour model.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr LB-111. doi:1538-7445.AM2012-LB-111