Figure 1.

Three-dimensional (3D) culture of prostate tumor spheroids in the absence or presence of bone marrow-derived adipocytes. (A) Schematic representation of 3D culture of tumor cells alone plated on reconstituted basement membrane (rBM)-coated coverslips with rBM overlay of 2% rBM and cultured alone (left) or in transwell with bone marrow adipocytes (right). (B) 3D reconstruction and DIC (differential interference contrast) images of the middle slice depicting morphology of the spheroids of PC3 cells grown for 3 (top) and 5 (bottom) days in control conditions or (C) in transwell coculture with adipocytes. 40× images; bar, 50 μm; nuclei are labeled with Hoechst dye (blue); green arrow (X), red arrow (Y), and blue arrow (Z) indicate orientation of the spheroid in 3D space. (D) Quantification of spheroid volume using Volocity software; data are shown in cubic micrometers and represent the mean (±SD) of three independent experiments with at least three independent spheroids measured/experiment; *p < 0.05 and ***p < 0.001 are considered statistically significant.