3D Oral Squamous Cell Carcinoma Microtissues Grown in Calcium Alginate Microbeads

Abstract

3D microtissue models, especially cancer microtissue model, are potentially applicable for new drug testing because 3D microtissue reveals more realistic drug response, characterizes the disease and mimics the tumor in human body. In this paper, we report the integration of oral squamous cell carcinoma (OSCC) cell line (ORL-48) into 3D microtissue after two weeks of microencapsulation in calcium alginate microbeads that were produced based on the flicking technique. The microtissues contained highly proliferative cells as indicated by Alamar blue assay. The viable microtissues formed were extracted from the calcium alginate shell by means of alginate lyase. As revealed by field emission scanning electron microscopy (FE-SEM), the extracted 3D microtissues were characterized by inhomogeneous microtissue surface but good cell integrity via self-secreted extracellular matrix proteins. DAPI staining showed the proliferative behavior of cells in multilayer structure. These microtissues were able to spread into 2D monolayer after being transferred to grow on petri dishes. The 3D cultured microtissues of ORL-48 could be potentially useful for cancer therapeutic drug assessment in-vitro.