Figure 1.

Schematic of process to obtain tumour-derived fibroblasts (TDF) from HNSCC patients and to incorporate the TDF into the lymphatic organotypic models. a) Schematic of the process to obtain and confirm phenotype of TDF from HNSCC patients. b) Microdevice picture and schematic (top- and cross-section view) showing TDFs within a collagen hydrogel on day 0. After collagen polymerization, the PDMS was removed to reveal a tubular structure that was lined with lymphatic endothelial cells (HLEC). c-d) Representative model staining. c) HLEC adhered in the lumen (red cell tracker) surrounded by HNSCC TDF (green cell tracker) from patient 1 at day 0. (Scale bar= 280 μm) d) Top-view of the lumen shows lumen confluency via F-actin (red) and nuclei (blue)(left) and 3D lumen structure (top-right). Prox-1 nuclear staining in red (lymphatic specific marker) and F-actin in green (bottom-right). (Scale bar= 140 μm).