Microtechnologies for Cell Microenvironment Control and Monitoring

Abstract

A great breadth of questions remains in cellular biology. Some questions cannot be answered using traditional analytical techniques and so demand the development of new tools for research. In the near future, the development of highly integrated microfluidic analytical platforms will enable the acquisition of unknown biological data. These microfluidic systems must allow cell culture under controlled microenvironment and high throughput analysis. For this purpose, the integration of a variable number of newly developed micro- and nano-technologies, which enable control of topography and surface chemistry, soluble factors, mechanical forces and cell–cell contacts, as well as technology for monitoring cell phenotype and genotype with high spatial and temporal resolution will be necessary. These multifunctional devices must be accompanied by appropriate data analysis and management of the expected large datasets generated. The knowledge gained with these platforms has the potential to improve predictive models of the behavior of cells, impacting directly in better therapies for disease treatment. In this review, we give an overview of the microtechnology toolbox available for the design of high throughput microfluidic platforms for cell analysis. We discuss current microtechnologies for cell microenvironment control, different methodologies to create large arrays of cellular systems and finally techniques for monitoring cells in microfluidic devices.

Recent Advances and Perspectives in Microfluidics‐Based Single‐Cell Biosensing Techniques

“…As an attempt to provide guidance for optimal selection and design of a suitable capture mechanism utilizing microfluidics for single‐cell analysis, this section briefly overviews the most commonly used microfluidic technologies applicable for cell sorting, isolation, and manipulation. These techniques include droplet encapsulation, flow cytometry (fluorescence‐activated cell sorting), antibody‐assisted capturing, micro‐array‐isolation, and field‐gradient‐based tweezers (such as optical, electrical, magnetic, acoustic, and hydrodynamic tweezers) …”

Continuous monitoring of cell transfection efficiency with micropatterned substrates

“…It constitutes the base of gene therapy, which aims to cure and treat genetic-related diseases and disorders through the insertion of specific genes or the manipulation and editing of existing, pathological genes (Kaufmann et al, 2013). Cell cultures are commonly used to carry out the transfection of cells and test, quantify and optimize transfection protocols that could lead to the development of efficient gene therapies (Azuaje-Hualde et al, 2017;Hamann et al, 2019).…”

“…Conventional cell culture lack control over the many cell-cell interactions and therefore, may not represent the best conditions for gene transfection. Moreover, it has been demonstrated that cell-substrate and cell-matrix interactions as well as cell architecture can affect cell's behavior and in turn affect cell transfection (Azuaje-Hualde et al, 2017;Mantz & Pannier, 2019). Over the past decade, several studies helped to better understand and control the interactions that may affect gene transfection.…”

Advances in Microtechnology for Improved Cytotoxicity Assessment

“…In the last decades, remarkable advancements have been made in the field of microtechnology to improve analytical processes in biology, through miniaturization, for biosensing DNA (Bulyk et al, 1999;Zhang et al, 2010) and protein arrays (He et al, 2008;Ramachandran et al, 2008;Lopez-Alonso et al, 2013;Gonzalez-Pujana et al, 2019), on-chip electrophoresis (Fritzsche et al, 2010;Ou, et al, 2019), microimmunoassays (Riahi et al, 2016;Hu et al, 2017), microfluidic cell sorting (Shields et al, 2015;Vaidyanathan et al, 2018) and for cellular membrane modelling (Hirano-Iwata et al, 2010;Strulson and Maurer, 2011;Galvez et al, 2020), among others (Beebe, et al, 2002;Sackmann, et al, 2014). In fact, microtechnology enables the precise control of the topography and the surface chemistry, leading to engineered platforms for the study of cellular processes or biosensing and, at the same time, bringing advantages such as time saving, reduced costs and working space, automation of the processes, increased sensitivity and reduced volumes of the required reagents (Wurm et al, 2010;Azuaje-Hualde et al, 2017).…”