title: A method for measurement of drug sensitivity of myeloma cells co-cultured with bone marrow stromal cells.
abstract: The tumor microenvironment can profoundly affect tumor cell survival as well as alter antitumor drug activity. However, conventional anticancer drug screening typically is performed in the absence of stromal cells. Here, we analyzed survival of myeloma cells co-cultured with bone marrow stromal cells (BMSC) using an automated fluorescence microscope platform, ScanR. By staining the cell nuclei with DRAQ5, we could distinguish between BMSC and myeloma cells, based on their staining intensity and nuclear shape. Using the apoptotic marker YO-PRO-1, the effects of drug treatment on the viability of the myeloma cells in the presence of stromal cells could be measured. The method does not require cell staining before incubation with drugs, and less than 5000 cells are required per condition. The method can be used for large-scale screening of anticancer drugs on primary myeloma cells. This study shows the importance of stromal cell support for primary myeloma cell survival in vitro, as half of the cell samples had a marked increase in their viability when cultured in the presence of BMSC. Stromal cell-induced protection against common myeloma drugs is also observed with this method.
authors: KG Jebsen Center for Myeloma Research and Department of Cancer Research and Molecular Medicine, Norwegian University of Science and Technology, Trondheim, Norway. kristine.misund@ntnu.no
authors: Kristine
authors: K
authors: Misund
authors: Katarzyna A
authors: KA
authors: Baranowska
authors: Toril
authors: T
authors: Holien
authors: Christoph
authors: C
authors: Rampa
authors: Dionne C G
authors: DC
authors: Klein
authors: Magne
authors: M
authors: Børset
authors: Anders
authors: A
authors: Waage
authors: Anders
authors: A
authors: Sundan
mesh_terms: D000970:Antineoplastic Agents / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects; D001583:Benzoxazoles / Q000378:metabolism; D002466:Cell Nucleolus / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D002470:Cell Survival / Q000187:drug effects; D018920:Coculture Techniques; D004354:Drug Screening Assays, Antitumor / Q000379:methods*; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D009101:Multiple Myeloma / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology; D011806:Quinolinium Compounds / Q000378:metabolism*; D014407:Tumor Cells, Cultured; D059016:Tumor Microenvironment / Q000187:drug effects
chemical_list: D000970:Antineoplastic Agents; D001583:Benzoxazoles; D011806:Quinolinium Compounds; C089813:YO-PRO 1
keywords: automated microscopy; bone marrow stromal cells; co-culture; drug sensitivity; multiple myeloma