Nanoliter scale microbioreactor array for quantitative cell biology

Abstract

A nanoliter scale microbioreactor array was designed for multiplexed quantitative cell biology. An addressable 8 Â 8 array of three nanoliter chambers was demonstrated for observing the serum response of HeLa human cancer cells in 64 parallel cultures. The individual culture unit was designed with a ''C'' shaped ring that effectively decoupled the central cell growth regions from the outer fluid transport channels. The chamber layout mimics physiological tissue conditions by implementing an outer channel for convective ''blood'' flow that feeds cells through diffusion into the low shear ''interstitial'' space. The 2 mm opening at the base of the ''C'' ring established a differential fluidic resistance up to 3 orders of magnitude greater than the fluid transport channel within a single mold microfluidic device. Three-dimensional (3D) finite element simulation were used to predict fluid transport properties based on chamber dimensions and verified experimentally. The microbioreactor array provided a continuous flow culture environment with a Peclet number (0.02) and shear stress (0.01 Pa) that approximated in vivo tissue conditions without limiting mass transport (10 s nutrient turnover). This microfluidic design overcomes the major problems encountered in multiplexing nanoliter culture environments by enabling uniform cell loading, eliminating shear, and pressure stresses on cultured cells, providing stable control of fluidic addressing, and permitting continuous on-chip optical monitoring. ß 2005 Wiley Periodicals, Inc.

Microfluidics-Based Cell Manipulation and Analysis

“…In this section, we mainly present the diversified cultivation works to date on both mammalian cells, bacteria, and plant cells using microfluidic devices. In the past 10 years, microfluidic-based applications of cell cultivation have ranged from many cell types from different tissues (e.g., epithelium and muscle) (Tourovskaia et al, 2005;Leclerc et al, 2006;Cimetta et al, 2009), organs (e.g., liver, lung, and kidney) (Zhang et al, 2008;Jang et al, 2010;Hoganson et al, 2011), even species (e.g., rat, cattle, human, and Nicotiana tabacum) (Ring et al, 2010;Taylor et al, 2005;Lee et al, 2006;Ko et al, 2006). Commonly, microfluidic culture modes of cells include the perfusion culture (continuous flow), half-perfusion culture (molecular diffusion with persistent supply of nutrients), and static culture (molecular diffusion).…”

Trends of Automation of Integrated Microfluidics

“…While silicon-based micro devices manufactured with methods developed in the semiconductor industry were some of the earliest examples of microfluidics-based research, there has been a shift over the last decade towards polymer-based devices, using materials such as Teflon, thermoset polyesters, silicon elastomer photoresist, SU-8 photoresist, poly-dimethylsiloxane (PDMS) and poly-methyl methacrylate [4][5][6][7]. Silicone rubber-based chemostats [8], bioreactors [9][10][11][12], and other microfluidic platforms [13,14] containing multiple cell chambers have been successfully applied in the cell culture applications in recent years. These microfluidic devices, consisting of optically-transparent PDMS, were fabricated using a casting process from silicon wafer molds containing photoresists with positive-relief channel patterns.…”

Lab-in-a-trench platform for real-time monitoring of cell surface protein expression

“…A range of concentrations can be readily established on the chip through the use of microfluidic gradient generators, which has motivated the construction of systems for the investigation of the cellular response to different reagent concentrations [14,15]. Multiple reagent concentrations have also been combined with multiple cell types on a single device to form twodimensional cell culture arrays for combinatorial analysis [16][17][18]. Additionally 2-dimensional cell culture arrays with different stimuli [19] and with different reporter cells [20] have also been demonstrated.…”