RESUMEN
Cell migration, which is regulated by intracellular signaling pathways (ICSP) and extracellular matrix (ECM), plays an indispensable role in many physiological and pathological process such as normal tissue development and cancer metastasis. However, there is a lack of rigorous and quantitative tools for analyzing the time-varying characteristics of cell migration in heterogeneous microenvironment, resulted from, e.g. the time-dependent local stiffness due to microstructural remodeling by migrating cells. Here, we develop a wavelet-analysis approach to derive the time-dependent motility parameters from cell migration trajectories, based on the time-varying persistent random walk model. In particular, the wavelet denoising and wavelet transform are employed to analyze migration velocities and obtain the wavelet power spectrum. Subsequently, the time-dependent motility parameters are derived via Lorentzian power spectrum. Our results based on synthetic data indicate the superiority of the method for estimating the intrinsic transient motility parameters, robust against a variety of stochastic noises. We also carry out a systematic parameter study and elaborate the effects of parameter selection on the performance of the method. Moreover, we demonstrate the utility of our approach via analyzing experimental data ofin vitrocell migration in distinct microenvironments, including the migration of MDA-MB-231 cells in confined micro-channel arrays and correlated migration of MCF-10A cells due to ECM-mediated mechanical coupling. Our analysis shows that our approach can be as a powerful tool to accurately derive the time-dependent motility parameters, and further analyze the time-dependent characteristics of cell migration regulated by complex microenvironment.
Asunto(s)
Movimiento Celular , Análisis de Ondículas , Línea Celular Tumoral , Células Epiteliales , HumanosRESUMEN
Increasingly being recognized is the role of the complex microenvironment to regulate cell phenotype; however, the cell culture systems used to study these effects in vitro are lagging. The complex microenvironment is host to a combination of biological interactions, chemical factors, and mechanical stimuli. Many devices have been designed to probe the effects of one mechanical stimulus, but few are capable of systematically interrogating all combinations of mechanical stimuli with independent control. To address this gap, we have developed the MechanoBioTester platform, a decoupled, multi-stimulus cell culture model for studying the cellular response to complex microenvironments in vitro. The system uses an engineered elastomeric chamber with a specially defined region for incorporating different target materials to act as the cell culture substrate. We have tested the system with several target materials including: polydimethylsiloxane elastomer, polyacrylamide gel, poly(1,8-octanediol citrate) elastomer, and type I collagen gel for both 2D and 3D co-culture. Additionally, when the chamber is connected to a flow circuit and our stretching device, stimuli in the form of fluid flow, cyclic stretch, and hydrostatic pressure are able to be imparted with independent control. We validated the device using experimental and computational methods to define a range of capabilities relevant to physiological microenvironments. The MechanoBioTester platform promises to function as a model system for mechanobiology, biomaterial design, and drug discovery applications that focus on probing the impact of a complex microenvironment in an in vitro setting. The protocol described within provides the details characterizing the MechanoBioTester system, the steps for fabricating the MechanoBioTester chamber, and the procedure for operating the MechanoBioTester system to stimulate cells.
Asunto(s)
Técnicas de Cultivo de Célula , Modelos Biológicos , Materiales Biocompatibles , BiofisicaRESUMEN
The objective of this research is to assess and analyze the biometeorological perception in complex microenvironments in the Athens University Campus (AUC) using urban micromodels, such as RayMan. The human thermal sensation in such a place was considered of great significance due to the great gathering of student body and staff of the University. The quantification of the biometeorological conditions was succeeded by the estimation of the physiologically equivalent temperature (PET), which is a biometeorological index based on the human energy balance. We carried out, on one hand, field measurements of air temperature, relative humidity, wind speed, and global solar irradiance for different sites (building atrium, open area, and green atrium) of the examined microurban environment in order to calculate PET during January-July 2013. Additionally, on the other hand, PET modeling was performed using different sky-view factors and was compared to a reference site (meteorological station of Laboratory of Climatology and Atmospheric Environment, University of Athens). The global radiation was transferred to the examined sites with the RayMan model, which considers the sky-view factors for the adaptation of the radiation fluxes to simple and complex environments. The results of this study reveal the crucial importance of the existence of trees and green cover in a complex environment, as a factor that could be the solution to the efforts of stake holders in order to mitigate strong heat stress and improve people's living quality in urban areas.