Figure 5.

Survival and functions of primary multiple myeloma (MM) cells and stroma are promoted in the scaffold. (A) Primary MM cells inside scaffold retain CD138 and light chain (κ chain) expression. (B) Western blot analyses performed on parallel 2D and 3D co-cultures of primary MM cells and MM bone marrow stromal cells (BMSC); pAkt, total Akt, β1 integrin (β1) and actin are depicted in a representative experiment (left) out of three. Right panels represent mean±Standard Error of Mean (SEM) of β1 integrin/actin and pAkt/total Akt ratios in three independent experiments. (C) Representative experiment of western blot analysis of STAT3 and Akt phosphorylation in 2D versus 3D co-cultures of primary MM cells and primary BMSC. Freshly isolated primary MM cells and BMSC are used as controls. (D) Input (t0) and recovered (t7) number of primary CD38⁺ MM cells from 7 patients after 3D culture for seven days. Dotted lines represent input and recovered number of CD38⁺ MM cells from 3 patients cultured in parallel 2D experiments. (E) Specialized functions of primary MM cells and stroma are measured in culture supernatants. Data are mean±SEM of six independent experiments. (F) IL-6 release in co-cultures with BMSC (left) and Ang-2 release in co-cultures with HUVEC (right) were determined by ELISA in parallel 2D and 3D experiments. Data are mean±SEM of three independent experiments. *P≤ 0.05; **P≤0.01. H&E: hematoxylin and eosin staining. Bar=100 mm.