In cancer research, either in diagnostics or anti-tumor drug development, it is important to fabricate a 3D tumor microenvironment in which all functions of cancer cells as metabolism, growth, migration and invasion could be achieved. Also, in drug development process, there is a huge need to fabricate high throughput 3D cell-based disease models that could be beneficial in toxicity testing and study of mechanism of drug effect. 3D cancer cell models as tumor spheroids, organoids, microfluidic based 3D cell aggregation and biomaterial-based 3D cell spheroids could make a proper mimic of real in vitro system. This research field has been grown during last years to develop and improve spheroid generation techniques. It is observed that cellulose family with their hydroxyl groups can interact with hydroxyl and carboxyl groups of collagen leading to spheroid formation which could be adjusted by α2β1 integrins on cancer cells which have an important role in cancer research. In this paper, we present a platform for spheroid generation using CNF and decellularized ECM (dECM) of liver tissue for 3D spheroid formation in anti-tumor drug screening. Layer-by-Layer (LbL) assembly containing CNF which was used as a method to coat microwell plates surfaces with varying parameters such as multiple cell seeding concentration, various tumor cell lines and a range of dECM concentration have been developed and characterized for spheroid formation and anticancer drug screening. The characterization methods with potency to characterize the LbL assembly of CNF were Atomic Force Microscopy (AFM) imaging and GISAXS measurements. Moreover, Bright field and Fluorescence images were provided to demonstrate the formation and viability of formed spheroids with and without dECM, before and after anticancer drug treatment.

Carboxymethylated cellulose nanofibrils (CNF): Application in 3D tumor modeling

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