A 3-D microfluidic combinatorial cell array

Abstract

We present the development of a three-dimensional (3-D) combinatorial cell culture array device featured with integrated three-input, eight-output combinatorial mixer and cell culture chambers. The device is designed for cell-based screening of multiple compounds simultaneously on a microfluidic platform. The final assembled device is composed of a porous membrane integrated in between a Parylene 3-D microfluidic chip and a PDMS microfluidic chip. The membrane turned the cell culture chambers into two-level configuration to facilitate cell loading and to maintain cells in a diffusion dominated space during device operation. Experimentally, we first characterized the combined compound concentration profile at each chamber using a fluorescence method. We then successfully demonstrated the functionality of the quantitative cell-based assay by culturing B35 rat neuronal cells on this device and screening the ability of three compounds (1,5-dihydroxyisoquinoline, deferoxamine, and 3-aminobenzoic acid) to attenuate cell death caused by cytotoxic hydrogen peroxide. In another experiment, we assayed for the combinatorial effects of three chemotherapeutic compound exposures (vinorelbine, paclitaxel, and γ-linolenic acid) on human breast cancer cells, MDA-MB-231. The same technology will enable the construction of inexpensive lab-on-a-chip devices with high-input combinatorial mixer for performing high-throughput cell-based assay and highly parallel and combinatorial chemical or biochemical reactions.

Microfluidic devices for cell cultivation and proliferation

“…Whilst in some cases some degree of stress has been found to induce growth and differentiation, 146 direct exposure to the flow rates generally required to supply sufficient nutrients is undesirable due to the damaging effect on the cells. Consequently, a wide range of approaches have been employed to protect cells from shear stress, including low flow rates, 43,48,102 changes in channel depth, 42 or geometry; 103,110 incorporation of microwells, 101 microchannels, 108,109 grooves, 41 micropillars, 66,67 or membranes 40,47,120,121 in or around the cell culture zone.…”

“…20 The diverse scope of microfluidics-based research in this area is evident from a survey of recent literature, which includes new approaches to precisely control fluid movement, [38][39][40] mimic biological functions, 40,41 study cell behaviour/interactions, [42][43][44][45] and undertake highthroughput cell culture and analysis. 39,46,47 3D cell cultures have been developed using gels as an extracellular matrix [48][49][50][51][52] or microstructural components to shape the 3D system. 53,54 …”

“…47 The device was composed of a Parylene C chip and a PDMS chip, with a porous polyethylene terephthalate (PET) membrane (0.4 or 1.0 lm pores) sandwiched in between the two chips. The bottom face of the Parylene chip contained the combinatorial mixer channels (14 lm height) and the top face contained the chambers, overpasses and outlet channels (25 lm height).…”

Fabrication of 3D high aspect ratio PDMS microfluidic networks with a hybrid stamp

“…3D microfluidics is one of the future trends since it not only significantly increases the number of functional units that can be integrated on a chip, thus increasing the processing power and throughput, but also provides many important unique functions that are difficult to realize with conventional 2D technologies, [2][3][4] such as 3D sheath flows for sample focusing 5,6 and 3D tissue engineering. [7][8][9][10][11][12][13][14] Versatile approaches have been demonstrated in the prior literature for fabricating 3D microfluidic structures. Siliconbased microfabrication allows the creation of high-resolution, high aspect ratio structures on silicon substrates, high conductivity metal electrodes for electrical sensing and actuation, and high quality semiconductor devices for IC integration.…”

A self-contained, programmable microfluidic cell culture system with real-time microscopy access

“…Among the advantages of microfluidics compared to conventional technologies are reduced sample consumption, portability, parallelization and increased spatiotemporal control of the sample environment (Liu et al 2010;Park et al 2010;Taylor et al 2009;Velve-Casquillas et al 2010) leading to possibilities for assays not possible with today's technology. Despite these advantages and the existence of highly advanced systems (Gomez-Sjöberg et al 2007;Taylor et al 2009) the microfluidic technology still has not made its entry as a common everyday tool into the biological research laboratories (Whitesides 2011;Young and Beebe 2010;Young and Simmons 2009).…”