Abstract
Multicellular spheroids have garnered significant attention as an in vitro three-dimensional cancer model which can mimick the in vivo microenvironmental features. While microfluidics generated double emulsions have become a potential method to generate spheroids, challenges remain on the tedious procedures. Enabled by a novel ‘airway resistance’ based selective surface treatment, this study presents an easy and facile generation of double emulsions for the initiation and cultivation of multicellular spheroids in a scaffold-free format. Combining with our previously developed DNA nanosensors, intestinal spheroids produced in the double emulsions have shown an elevated activities of an essential DNA modifying enzyme, the topoisomerase I. The observed molecular and functional characteristics of spheroids produced in double emulsions are similar to the counterparts produced by the commercially available ultra-low attachment plates. However, the double emulsions excel for their improved uniformity, and the consistency of the results obtained by subsequent analysis of the spheroids. The presented technique is expected to ease the burden of producing spheroids and to promote the spheroids model for cancer or stem cell study.
Highlights
Mounting evidence has suggested that the cell-to-cell interactions and extracellular matrix (ECM) mechanics are distinctively different, when cells are interacting in a threedimensional (3D) architecture as opposed to in conventional two-dimensional (2D) cell cultures [1]
To satisfy the request of facile surface treatment of PDMS selectively, this study proposes a novel design, which is composed of two cross-junctions and a serpentine structure in between, as shown in figure 1(a)
As a continuous advancement of our previous effort, this study presents a novel approach to selectively modify the wettability of PDMS, by the introduction of airway resistance
Summary
Mounting evidence has suggested that the cell-to-cell interactions and extracellular matrix (ECM) mechanics are distinctively different, when cells are interacting in a threedimensional (3D) architecture as opposed to in conventional two-dimensional (2D) cell cultures [1]. In accordance with the recognition that 2D cell cultures lack the adequate cell–cell interactions and ECM presented in the human tissues, 3D cell culture models have gained growing interest for the studies of tumours and tissue engineering due to their obvious advantages of mimicking the in vivo environments of human cells With this being said, proper 3D architectures are important to the cell types presented in the connective tissues or sensitive to the mechanical stresses, such as fibroblasts, epithelium, and cancer cells [8]. Previous studies have shown that stimulation of the ECM in 3D culture with selective incorporation of adhesion factors and signalling factors is capable of turning the epithelia carcinoma cells into non-cancerous cells [4, 10,11,12] Such cells have shown a relatively high degree of drug resistance compared to their counterparts cultured in conventional 2D models. Xenograft tumour sections from the nude mice and tumour spheroids appear to have similar hematoxylin and eosin staining [17]
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