Live-Cell Systems in Real-Time Biomonitoring of Water Pollution: Practical Considerations and Future Perspectives

Continuous monitoring and early warning of potential water contamination with toxic chemicals is of paramount importance for human health and sustainable food production. During the last few decades there have been noteworthy advances in technologies for the automated sensing of physicochemical parameters of water. These do not translate well into online monitoring of chemical pollutants since most of them are either incapable of real-time detection or unable to detect impacts on biological organisms. As a result, biological early warning systems have been proposed to supplement conventional water quality test strategies. Such systems can continuously evaluate physiological parameters of suitable aquatic species and alert the user to the presence of toxicants. In this regard, single cellular organisms, such as bacteria, cyanobacteria, micro-algae and vertebrate cell lines, offer promising avenues for development of water biosensors. Historically, only a handful of systems utilising single-cell organisms have been deployed as established online water biomonitoring tools. Recent advances in recombinant microorganisms, cell immobilisation techniques, live-cell microarrays and microfluidic Lab-on-a-Chip technologies open new avenues to develop miniaturised systems capable of detecting a broad range of water contaminants. In experimental settings, they have been shown as sensitive and rapid biosensors with capabilities to detect traces of contaminants. In this work, we critically review the recent advances and practical prospects of biological early warning systems based on live-cell biosensors. We demonstrate historical deployment successes, technological innovations, as well as current challenges for the broader deployment of live-cell biosensors in the monitoring of water quality.

Stem Cell Microarrays for Assessing Growth Factor Signaling in Engineered Glycan Microenvironments

The incorporation of extracellular matrix (ECM) elements such as adhesion motifs, growth factors and signaling molecules provides unique functionalities to biomaterials in regenerative medicine. However, an often overlooked component of native microenvironments are extracellular glycans, which are key players in mediating biological processes in signaling and development. Here, we describe a cellular microarray which presents well-defined glycan structures in a synthetic hydrogel ECM microenvironment to colonies of embryonic stem cells (ESCs) in a spaitally adressable format. In conjunction with synthetic glycan microenvironments, this new platform enables screening well-defined desulfated heparin glycans in the extracellular space by linking protein-glycan interactions directly with mitogen activated protein kinase (MAPK) signaling events leading to differentiation. The cellular array technique provides a potent tool to rapidly optimize the glycan ECM and providing guiding principles for incorporation of functional glycan elements into biomaterial design.

Robotic Cell Printing for Constructing Living Yeast Cell Microarrays in Microfluidic Chips

Living cell microarrays in microfluidic chips allow the non-invasive multiplexed molecular analysis of single cells. Here, we developed a simple and affordable perfusion microfluidic chip containing a living yeast cell array composed of a population of cell variants (green fluorescent protein (GFP)-tagged Saccharomyces cerevisiae clones). We combined mechanical patterning in 102 microwells and robotic piezoelectric cell dispensing in the microwells to construct the cell arrays. Robotic yeast cell dispensing of a yeast collection from a multiwell plate to the microfluidic chip microwells was optimized. The developed microfluidic chip and procedure were validated by observing the growth of GFP-tagged yeast clones that are linked to the cell cycle by time-lapse fluorescence microscopy over a few generations. The developed microfluidic technology has the potential to be easily upscaled to a high-density cell array allowing us to perform dynamic proteomics and localizomics experiments.