It is known that cellCECM interactions affects cellCcell tension, and therefore, cellCcell contact, resulting in an interdependence of cellCcell and cellCECM interactions . 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. strong class=”kwd-title” Keywords: cell analysis, high-throughput, microfluidics, microtechnology 1. Introduction Native cells are in a dynamic multifactorial environment, their own microenvironment. The cell microenvironment is constituted by: their extracellular matrix (ECM), the topography and physical properties of the ECM and by soluble factors on their fluidic environment. All of them strongly affect cell fate and cell behavior. Changes in the cell microenvironment are transduced into intracellular signaling pathways, which regulate cell fate and cell behavior. Conventional cell culture systems often rely on batch experiments with limited control of cell microenvironments. In order to obtain a comprehensive knowledge of cell function and behavior, it would be desirable to develop experimental methods that could explain the contribution of each of those environmental factors, as well as their synergetic effects on cell behavior (Figure 1). Open in a separate window Figure 1 Input signals from cell microenvironment induce internal signaling of cells and modulate their outputs, affecting cell behavior. During the last two decades, we have witnessed a number of key developments in the area of the microtechnologies, which allows introducing control and complexity over a full range of environmental factor at the microscale level. For example, technologies for the accurate structuration of surfaces for subsequent cell culture, microfluidic architectures, synthesis of novel biomaterials and nanomaterials with sensing and actuating capabilities have been developed and their potential for cell culture, stimulation and analysis has been proven. In particular, the miniaturized scale of microchannels in microfluidic devices offers advantages such as low contamination risk, fast transfer of nutrients and heat, short equilibration times, parallelization of processes and automation, low reagent and power consumption, portability, etc. Moreover, since the dimensional environment is analogous to in vivo conditions, the small sizes of the channels permit medium and nutrients to diffuse to nutrient-poor areas. Currently, there is little development of microtechnologies that can successfully mimic the in vivo microenvironments, since any change in material, surface chemistry, cell number or flow conditions can affect the results of the assays . Nowadays, there is Gabapentin enacarbil an increasing use of microfluidic techniques on cell culture that have opened a broad range of possibilities for studying cells in a variety of contexts, allowing to understand the specific contribution of each different parameter to cellular behavior, such as shear forces, nutrient gradients, etc. . An extra advantage of the use of microtechnologies is the scalability and the possibility of parallelization of cellular samples which allow high-throughput (HTP) measurements, essential for the Gabapentin enacarbil statistical analysis of multi-parameter environments, and for the construction of predictive models. The current trend is to develop HTP and multiplexed technologies, essentially those who also allow a real time or near-real time analysis for both single cell and multi cell platforms. The properties that can be quantified from analysis includes the study of the cells mechanics (deformation, migration and growth), the proteome, genome and secretome, and both their extracellular and intracellular interactions and their stimuli . Integration of several microtechnologies to create controlled multi-parametric environments and monitoring is still a challenge. Microfluidics has emerged as a new way to fabricate large cellular arrays in defined patterns which allows Gabapentin enacarbil the study of a Dll4 large number of cells in a specific microenvironment as well as the observation and quantification of several outcomes from a single study. Looking for the best way to design novel platforms for cell analysis, in this manuscript, we review examples on how different parameters of cell microenvironment may be controlled through microtechnologies, as well as the techniques available for monitoring Gabapentin enacarbil cells in microfluidic devices, centering on the analysis of chemicals outcomes. Additionally, we give an overview of current microfluidic platforms already available for cell analysis. 2. Microtechnologies for Cell Microenvironment Control The term cell microenvironment involves those factors that directly or indirectly affect cell behavior, by biophysical, biochemical or other pathways. As previously stated, those factors can be classified in: ECM, cells surrounding a single cell, soluble factors, topography or physical properties of the ECM, and fluidic cues. All of them strongly affect cell proliferation and differentiation. However, despite this distinction, each factor cannot be considered independently, as all affect cell.