A Method for Measurement of Drug Sensitivity of Myeloma Cells Co-Cultured with Bone Marrow Stromal Cells

摘要: Abstract 1373 The aim of this work was to establish a robust and simple method for the measurement drug sensitivity in myeloma cells under conditions mimicking aspects bone marrow microenvironment. In particular we wanted measure cultivated presence stromal cells. The tumor microenvironment can profoundly affect cell survival as well alter antitumor activity, it is generally believed that growth critically dependent on Bone (BMSC) have been shown protect from common cytostatic or cytotoxic drugs vitro. Common vitro assays used high-throughput screening cannot easily discriminate between responses co-cultures. Although few recent studies overcome problem (Ramasamy K. et al. , 157(5):564–79,2012, McMillin D. 16(4):483–9, 2010), application stable transfection labeling limits practical these co-culture lines, excluding primary inherently may be hard transduce even by retroviral vectors. Here, analyzed co-cultured with BMSC using an automated fluorescence microscope, ScanR. ScanR microscope based station. By staining nuclei DRAQ5, were able cells, their intensity nuclear shape. Using apoptotic marker YO-PRO-1, effects treatment viability could measured. The main advantages are non-necessity manipulation before low number (5000 cells) needed per measurement, which makes ideal experiments cells. fact, analysis easier more when slowly growing i.e . compared rapidly proliferating lines. This should well-suited high throughput analysis, stained situ no washing, centrifugation, fixation steps analysis. The conventional detecting viability; flow cytometry where annexin V detect As [figure 1][1], dose-response curves obtained ANBL-6 treated different doses melphalan similar showed same trends both methods. However, evident system than (EC50 YO-PRO-1 = 11μM versus EC50Annexin V= 15μM). The population applied study support IL-6 lines without addition IL-6. INA-6 survived absence added shows importance half samples had marked increase cultured BMSC. Stromal cell-induced protection against also observed method. For instance, patient MM7, BMSC, EC50 cyclophosphamide increased 5 μM approximately 10 ([figure 2][2]). ![Figure 1][3] Figure 1 ![Figure 2][3] 2 Disclosures: No relevant conflicts interest declare. [1]: #F1 [2]: #F2 [3]: pending:yes