Abstract
We present a simple bench-top method to fabricate enclosed circular channels for biological experiments. Fabricating the channels takes less than 2 hours by using glass capillaries of various diameters (from 100 µm up to 400 µm) as a mould in PDMS. The inner surface of microchannels prepared in this way was coated with a thin membrane of either Matrigel or a layer-by-layer polyelectrolyte to control cellular adhesion. The microchannels were then used as scaffolds for 3D-confined epithelial cell culture. To show that our device can be used with several epithelial cell types from exocrine glandular tissues, we performed our biological studies on adherent epithelial prostate cells (non-malignant RWPE-1 and invasive PC3) and also on breast (non-malignant MCF10A) cells We observed that in static conditions cells adhere and proliferate to form a confluent layer in channels of 150 µm in diameter and larger, whereas cellular viability decreases with decreasing diameter of the channel. Matrigel and PSS (poly (sodium 4-styrenesulphonate)) promote cell adhesion, whereas the cell proliferation rate was reduced on the PAH (poly (allylamine hydrochloride))-terminated surface. Moreover infusing channels with a continuous flow did not induce any cellular detachment. Our system is designed to simply grow cells in a microchannel structure and could be easily fabricated in any biological laboratory. It offers opportunities to grow epithelial cells that support the formation of a light. This system could be eventually used, for example, to collect cellular secretions, or study cell responses to graduated hypoxia conditions, to chemicals (drugs, siRNA, …) and/or physiological shear stress.
Highlights
Model systems that recreate the architectural features observed in tissues and in tumours are of prime interest to study morphogenesis and carcinogenesis
Our motivation for this work was to develop polyelectrolyte 3D scaffolds in order to create new cell-culture devices that more closely mimic the natural physiological ductal environment found in exocrine glands
The present work described in this manuscript provides a detailed protocol for bench-top rapid fabrication of circular channels for 3D cell culture (Figure 1 & Figure S1 in File S1)
Summary
Model systems that recreate the architectural features observed in tissues and in tumours are of prime interest to study morphogenesis and carcinogenesis. Numerous approaches have emerged that are aimed to control the spatial and temporal properties of the cell microenvironment (e.g. stiffness, 3D structure, micropatterning, shear stress). Endothelial cells cultured within a chamber capable of applying physiological shear stresses are induced to differentiate due to stimulation of specific integrin/endothelial cell-mediated signalling cascades [2]. Epithelial cells cultured on soft extracellular matrix gels organize themselves into polarized structures that strongly resemble functional tissue in vivo [3,4]. For in vitro studies of vascular tissue, lab-on-chip (LOC) systems [5] that incorporate a unique 3D and dynamic microenvironment with high spatiotemporal precision provide a physiologically relevant way to reproduce vascular tissues
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