Planar Gradient Diffusion System to Investigate Chemotaxis in a 3D Collagen Matrix

Abstract

The importance of cell migration can be seen through the development of human life. When cells migrate, they generate forces and transfer these forces to their surrounding area, leading to cell movement and migration. In order to understand the mechanisms that can alter and/or affect cell migration, one can study these forces. In theory, understanding the fundamental mechanisms and forces underlying cell migration holds the promise of effective approaches for treating diseases and promoting cellular transplantation. Unfortunately, modern chemotaxis chambers that have been developed are usually restricted to two dimensions (2D) and have complex diffusion gradients that make the experiment difficult to interpret. To this end, we have developed, and describe in this paper, a direct-viewing chamber for chemotaxis studies, which allows one to overcome modern chemotaxis chamber obstacles able to measure cell forces and specific concentration within the chamber in a 3D environment to study cell 3D migration. More compelling, this approach allows one to successfully model diffusion through 3D collagen matrices and calculate the coefficient of diffusion of a chemoattractant through multiple different concentrations of collagen, while keeping the system simple and user friendly for traction force microscopy (TFM) and digital volume correlation (DVC) analysis. Video LinkThe video component of this article can be found at

The Immunobiogram, a Novel In Vitro Assay to Evaluate Treatment Resistance in Patients Receiving Immunosuppressive Therapy

“…Spontaneous diffusion of a substance through hydrogel depends on several parameters, including the characteristics of the solute (e.g., concentration, molecular size, and solubility, polarity) and of the hydrogel network (e.g., pore size, chemical monomer structure, crosslink degree, and mesh size), temperature, pH, and the presence of ions ( 25 ). Upon placement of the immunosuppressant disc at one end of a channel on the IMBG plate, the immunosuppressant contained in the disc diffuses passively through the hydrogel in the channel, producing a stable and quantifiable drug concentration gradient ( 26 , 27 ). This drug concentration gradient exerts a dose-dependent inhibitory effect on the activation and proliferation capacity of the PBMCs embedded in the hydrogel.…”

Neutrophil Recruitment: From Model Systems to Tissue-Specific Patterns

“…Thus, these findings together with migration experiments in integrin-depleted murine leukocytes, suggest that integrin-dependent and -independent processes can differentially manifest in 2D vs. 3D migratory environments [31]. Aside from commerciallyavailable chambers utilizing transmigration-or collagen-based chemotaxis assays [32], a planar gradient system was recently developed to image neutrophil chemotaxis in 3D within a collagen matrix; using this system, the precise positioning and force generation of cells in the 3D z-direction (in addition to the x and y axes) could be assessed [33]. This was accomplished by preparation of a lightpenetrable collagen-medium, in which chemokine diffusion could be measured, accessible by confocal microscopy in combination with traction-force microscopy.…”

Identification and Characterisation of tracheal cartilage derived stem cells for airway tissue engineering

“…Cultispher® microcarriers encouraged cells to undergo chondrogenesis the large spaces in between each individual microcarriers in all four conditions mean lack of cell-cell contact which is one of the main key factors in cartilage formation [154]. Moreover, the matrix deposition exclusively occurred on the periphery of the constructs in all four groups and the lack of nutrient supply to the center of constructs might have caused cell death or chemotaxis of cells [470] to the periphery where a layer of viable cells was clearly visible. To improve mechanical strength and shape fidelity the use of bioprinting and other co-polymers may be further investigated.…”

“…The lack of matrix synthesis in the centre and middle part of disks could be explained by lack of cell-cell contact, which is a prerequisite for cartilage condensation and formation [154]. Lack of cellular content in the centre of the disk may also be due to chemotaxis of cells [470] to the surface of the disk for the purpose of access to nutrients or cellular necrosis due to reduced oxygen and nutrients in the centre of the disks. One possibility is that the periphery of constructs had formed matrix around the edges and possible lack of diffusion in nutrients to the inner depth of disks could have impacted cells to migrate towards the periphery [470].…”

“…Lack of cellular content in the centre of the disk may also be due to chemotaxis of cells [470] to the surface of the disk for the purpose of access to nutrients or cellular necrosis due to reduced oxygen and nutrients in the centre of the disks. One possibility is that the periphery of constructs had formed matrix around the edges and possible lack of diffusion in nutrients to the inner depth of disks could have impacted cells to migrate towards the periphery [470]. Using blood derived biological glues have shown to improve disk formation when coupled with human articular chondrocytes and microcarriers, and this effect may have been due…”