RESUMEN
The mechanical cue of fiber alignment plays a key role in the development of various tissues in the body. The ability to study the effect of these stimuli in vitro has been limited previously. Here, we present a microfluidic device capable of intrinsically generating aligned fibers using the microchannel geometry. The device also features tunable interstitial fluid flow and the ability to form a morphogen gradient. These aspects allow for the modeling of complex tissues and to differentiate cell response to different stimuli. To demonstrate the abilities of our device, we incorporated luminal epithelial cysts into our device and induced growth factor stimulation. We found the mechanical cue of fiber alignment to play a dominant role in cell elongation and the ability to form protrusions was dependent on cadherin-3. Together, this work serves as a springboard for future potential with these devices to answer questions in developmental biology and complex diseases such as cancers.
Asunto(s)
Morfogénesis , Animales , Quimiocinas/metabolismo , Microfluídica/métodos , Células Epiteliales/metabolismo , Células Epiteliales/citología , Cadherinas/metabolismo , Dispositivos Laboratorio en un Chip , Epitelio/metabolismo , Epitelio/crecimiento & desarrollo , Perros , Humanos , Matriz Extracelular/metabolismo , Matriz Extracelular/química , Células de Riñón Canino Madin Darby , Modelos BiológicosRESUMEN
Leader cells direct collective migration through sensing cues in their microenvironment to determine migration direction. The mechanism by which leader cells sense the mechanical cue of organized matrix architecture culminating in a mechanical response is not well defined. In this study, we investigated the effect of organized collagen matrix fibers on leader cell mechanics and demonstrate that leader cells protrude along aligned fibers resulting in an elongated phenotype of the entire cluster. Further, leader cells show increased mechanical interactions with their nearby matrix compared to follower cells, as evidenced by increased traction forces, increased and larger focal adhesions, and increased expression of integrin-α2. Together our results demonstrate changes in mechanical matrix cues drives changes in leader cell mechanoresponse that is required for directional collective migration. Our findings provide new insights into two fundamental components of carcinogenesis, namely invasion and metastasis.
Asunto(s)
Colágeno , Movimiento Celular , Colágeno/farmacologíaRESUMEN
A fundamental step of tumor metastasis is tumor cell migration away from the primary tumor site. One mode of migration that is essential but still understudied is collective invasion, the process by which clusters of cells move in a coordinated fashion. In recent years, there has been growing interest to understand factors regulating collective invasion, with increasing number of studies investigating the biomechanical regulation of collective invasion. In this review we discuss the dynamic relationship between tumor microenvironment cues and cell response by first covering mechanical factors in the microenvironment and second, discussing the mechanosensing pathways utilized by cells in collective clusters to dynamically respond to mechanical matrix cues. Finally, we discuss model systems that have been developed which have increased our understanding of the mechanical factors contributing to tumor progression.
Asunto(s)
Modelos Biológicos , Neoplasias , Humanos , Microambiente TumoralRESUMEN
Non-specific adsorption (NSA) is a persistent problem that negatively affects biosensors, decreasing sensitivity, specificity, and reproducibility. Passive and active removal methods exist to remedy this issue, by coating the surface or generating surface forces to shear away weakly adhered biomolecules, respectively. However, many surface coatings are not compatible or effective for sensing, and thus active removal methods have been developed to combat this phenomenon. This review aims to provide an overview of methods of NSA reduction in biosensing, focusing on the shift from passive methods to active methods in the past decade. Attention is focused on protein NSA, due to their common use in biosensing for biomarker diagnostics. To our knowledge, this is the first review to comprehensively discuss active NSA removal methods. Lastly, the challenges and future perspectives of NSA reduction in biosensing are discussed.