January 11, 2018
Then, 21 concentrations of afatinib, gefitinib, pictilisib, GDC-0068, AZD8055, everolimus (all Selleckchem), or tamoxifen (Sigma) as well as DMSO controls were added in duplicate using a Tecan D300e digital dispenser (Life Sciences). To measure ATP as proxy for viable cells, 40 ÎźL of CellTiter-Glo 3D Reagent (Promega) per well were added after five days, before shaking the plate for 30 min at room temperature and reading luminescence on a SpectraMax microplate reader (Molecular Devices)
Breast cancer (BC) comprises multiple distinct subtypes that differ genetically, pathologically, and clinically. Here, we describe a robust protocol for long-term culturing of human mammary epithelial organoids. Using this protocol, >100 primary and metastatic BC organoid lines were generated, broadly recapitulating the diversity of the disease. BC organoid morphologies typically matched the histopathology, hormone receptor status, and HER2 status of the original tumor. DNA copy number variations as well as sequence changes were consistent within tumor-organoid pairs and largely retained even after extended passaging. BC organoids furthermore populated all major gene-expression-based classification groups and allowed in vitro drug screens that were consistent with in vivo xeno-transplantations and patient response. This study describes a representative collection of well-characterized BC organoids available for cancer research and drug development, as well as a strategy to assess in vitro drug response in a personalized fashion. Copyright Š 2017 Elsevier Inc. All rights reserved.
Sachs, N; de Ligt, J; Kopper, O; Gogola, E; Bounova, G; Weeber, F; Balgobind, AV; Wind, K; Gracanin, A; Begthel, H; Korving, J; van Boxtel, R; Duarte, AA; Lelieveld, D; van Hoeck, A; Ernst, RF; Blokzijl, F; Nijman, IJ; Hoogstraat, M; van de Ven, M; Egan, DA; Zinzalla, V; Moll, J; Boj, SF; Voest, EE; Wessels, L; van Diest, PJ; Rottenberg, S; Vries, RGJ; Cuppen, E; Clevers, H;
Journal: Cell Pages: 373-386.e10