In the past decade, it has been well recognised that the tumour microenvironment contains microenvironmental components such as
hypoxia that significantly influence tumour cell behaviours such, invasiveness and
therapy resistance, all of which provide new targets for studying
cancer biology and developing anticancer
therapeutics. In response, a large number of two-dimensional and three-dimensional (3D) in vitro tumour models have been developed to recapitulate different aspects of the tumour microenvironment and enable the study of related biological questions. While more complex models enable new biological insight, such models often involve time-consuming and complex fabrication or analysis processes, which limit their adoption by the broader
cancer biology community. To address this, we recently reported the development of a new platform that enables easy assembly and analysis of 3D tumour cultures, the tissue roll for analysis of cellular environment response (TRACER). The TRACER platform enables recapitulation of many spatial aspects of the tumour microenvironment to ask a variety of questions, however its original design contains only one cell type. In contrast tumours in vivo often contain a neoplastic and stromal compartment. To expand the types of questions the TRACER system is useful for asking, here we present a strategy to pattern distinct cell type domains into TRACER layers using a custom-built
gelatin-dispensing pen. The pen allows deposition of a temporary
gelatin barrier into the TRACER scaffold to define domain boundaries between cell populations. The
gelatin can be melted away after cell seeding to allow interaction of cell populations from adjacent domains. Our device offers a simple strategy to generate complex multi-cell type tumour cultures for analysis of fundamental biology and drug development applications.