May 1, 2019
The drugs and their combinations were added 1 h after plating the organoids using the Tecan D300e Digital Dispenser. Drugs were dispensed in a randomized manner and DMSO end concentration was 1% in all wells. 120 h after adding the drugs, ATP levels were measured using the Cell-Titer Glo2.0 (Promega BV) according to the manufacturerâs instructions and luminescence was measured using a SpectraMax microplate reader (Molecular Devices). Results were normalized to vehicle (DMSO = 100%) and baseline control (navitoclax 20 ÂľM)
Ovarian cancer (OC) is a heterogeneous disease usually diagnosed at a late stage. Experimental in vitro models that faithfully capture the hallmarks and tumor heterogeneity of OC are limited and hard to establish. We present a protocol that enables efficient derivation and long-term expansion of OC organoids. Utilizing this protocol, we have established 56 organoid lines from 32 patients, representing all main subtypes of OC. OC organoids recapitulate histological and genomic features of the pertinent lesion from which they were derived, illustrating intra- and interpatient heterogeneity, and can be genetically modified. We show that OC organoids can be used for drug-screening assays and capture different tumor subtype responses to the gold standard platinum-based chemotherapy, including acquisition of chemoresistance in recurrent disease. Finally, OC organoids can be xenografted, enabling in vivo drug-sensitivity assays. Taken together, this demonstrates their potential application for research and personalized medicine.
Kopper, O; de Witte, CJ; LĂľhmussaar, K; Valle-Inclan, JE; Hami, N; Kester, L; Balgobind, AV; Korving, J; Proost, N; Begthel, H; van Wijk, LM; Revilla, SA; Theeuwsen, R; van de Ven, M; van Roosmalen, MJ; Ponsioen, B; Ho, VWH; Neel, BG; Bosse, T; Gaarenstroom, KN; Vrieling, H; Vreeswijk, MPG; van Diest, PJ; Witteveen, PO; Jonges, T; Bos, JL; van Oudenaarden, A; Zweemer, RP; Snippert, HJG; Kloosterman, WP; Clevers, H;
Journal: Nat. Med. Pages: 838-849