Figure 3.

Decellularized lymph node matrices repopulated with lymph nodemesenchymal stromal and Hodgkin lymphoma cells as 3D culture model to test ADAM10 inhibitors. (A and B) Immunohistochemistry (IHC) of 3D cultures performed on decellularized Hodgkin lymphoma (HL) lymph node (LN) matrices (extracellular matrix [ECM], one reprentative experiment, LN I-19032-16) with 2x10⁵ LN-MSC16412 for 3 days, followed by the addition of 4x10⁵ L428 cells to each ECM/LNMSC16412 scaffold for a further 2 days. (A) 4 mm sections stained with the anti-CD30 monoclonal antibody (mAb), to identify HL (L428) cells, followed by Biot-GAM, HRP-Av and developed with DAB; (B) sections stained with the anti- TGII rabbit polyclonal antiserum, to identify LN-MSC16412, followed by Biot- GAR, HRP-Av and developed with DAB. Inset in (A) negative control with Biot- GAM alone. Slides were counterstained with hematoxylin and analyzed under a Leica DM MB2 microscope (40x objective). (C to D) TNFa C) or soluble CD30 (D) content (pg/mL/10⁶ cells) in the supernatant (SN) recovered after 48 hour (h) from the addition of 10 mM LT4 or MN8 or GIX ADAM10 inhibitors to the 3D cultures, measured by specific enzyme-linked immunosorbant assay. Nil: solvent (dimethylsulfoxide 1:1,000). Results are the mean ± standard deviation from three experiments performed in duplicate with ECM from three HL patients.*P<0.005 vs. nil; **P<0.001 vs. nil.