Generating 3D artificial melanoma tumor tissues using CellMate™ hydrogels.

Abstract

We have previously reported on the successful use of natural 3D ECM materials as artificial tissue scaffolds to generate tissue‐like constructs of melanoma using the ATCC cell lines B16F1and B16F10. In that earlier work we observed that 3D constructs like these undergo a progressive scaffold colonization, tissue development and maturation in‐vitro which closely parallels tumor progression in‐vivo. Now, we have extended that work by applying a new and unique 3D hydrogel matrix called CellMate™. This material is an embedding hydrogel comprised of hyaluronate and chitosan which crosslink to generate a matrix scaffolding for cell attachment and tissue development. In these new experiments, B16F1 and B16F10 cell lines were each seeded into CellMate™ at densities of 10 and 20 million cells per scaffold. The resulting structures where then maintained for 6 months of continuous culture with periodic new plates and regular media exchanges at 3 day intervals. This approach resulted in even more physiological artificial tumors, than previously developed on ECM scaffolds. As previously seen on natural scaffolds, these new artificial tumors develop two distinct cell populations which closely reflect the normal in‐vivo tumor structures seen in the clinical setting. These two cell types include pigmented cells with a relatively dendritic appearance and non‐pigmented cells with relatively less dendritic and more rounded cell bodies. These cellular phenotypes also gave rise to distinct tissue‐level structural phenotypes as well. Pigmented tissue regions tended to present with less regular surfaces and more tendency to produce branching structures that extend away from the construct. In contrast, non‐pigmented cell regions generated large rounded and bulging features which often resembled ridges of tissue and which had very smooth surface features. Additionally these structures also appeared to be highly friable and prone to peeling off the main pigmented tumor structure. Colonies and later cell‐strains were developed from these shed structures and these retained their non‐pigmented phenotypes through multiple passages in 2D monolayers. Collectively these studies illustrate the efficacy of using 3D artificial tissues to create better in‐vitro tumor models of melanoma.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.