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Spatial patterning of different cell types is crucial for tissue engineering and is characterized by the formation of sharp boundary between segregated groups of cells of different lineages. The cell−cell boundary layers, depending on the relative adhesion forces, can result in kinks in the border, similar to fingering patterns between two viscous partially miscible fluids which can be characterized by its fractal dimension. This suggests that mathematical models used to analyze the fingering patterns can be applied to cell migration data as a metric for intercellular adhesion forces. In this study, we develop a novel computational analysis method to characterize the interactions between blood endothelial cells (BECs) and lymphatic endothelial cells (LECs), which form segregated vasculature by recognizing each other through podoplanin. We observed indiscriminate mixing with LEC−LEC and BEC−BEC pairs and a sharp boundary between LEC−BEC pair, and fingering-like patterns with pseudo-LEC−BEC pairs. We found that the box counting method yields fractal dimension between 1 for sharp boundaries and 1.3 for indiscriminate mixing, and intermediate values for fingering-like boundaries. We further verify that these results are due to differential affinity by performing random walk simulations with differential attraction to nearby cells and generate similar migration pattern, confirming that higher differential attraction between different cell types result in lower fractal dimensions. We estimate the characteristic velocity and interfacial tension for our simulated and experimental data to show that the fractal dimension negatively correlates with capillary number (Ca), further indicating that the mathematical models used to study viscous fingering pattern can be used to characterize cell−cell mixing. Taken together, these results indicate that the fractal analysis of segregation boundaries can be used as a simple metric to estimate relative cell−cell adhesion forces between different cell types.

 

Encapsulation of single cells in a thin hydrogel provides a more precise control of stem cell niches and better molecular transport. Despite the recent advances in microfluidic technologies to allow encapsulation of single cells, existing methods rely on special crosslinking agents that are pre-coated on the cell surface and subject to the variation of the cell membrane, which limits their widespread adoption. This work reports a high-throughput single-cell encapsulation method based on the “tip streaming” mode of alternating current (AC) electrospray, with encapsulation efficiencies over 80% after tuned centrifugation. Dripping with multiple cells is curtailed due to gating by the sharp conic meniscus of the tip streaming mode that only allows one cell to be ejected at a time. Moreover, the method can be universally applied to both natural and synthetic hydrogels, as well as various cell types, including human multipotent mesenchymal stromal cells (hMSCs). Encapsulated hMSCs maintain good cell viability over an extended culture period and exhibit robust differentiation potential into osteoblasts and adipocytes. Collectively, electrically induced tip streaming enables high-throughput encapsulation of single cells with high efficiency and universality, which is applicable for various applications in cell therapy, pharmacokinetic studies, and regenerative medicine. 

In this study, an ion depleted zone created by an ion-selective membrane was used to impose a high and uniform constant extracellular potential over an entire ∼1000 cell rat cardiomyocyte (rCM) colony on-a-chip to trigger synchronized voltage-gated ion channel activities while preserving cell viability, thus extending single-cell voltage-clamp ion channel studies to an entire normalized colony. Image analysis indicated that rCM beating was strengthened and accelerated (by a factor of ∼2) within minutes of ion depletion and the duration of contraction and relaxation phases was significantly reduced. After the initial synchronization, the entire colony responds collectively to external potential changes such that beating over the entire colony can be activated or deactivated within 0.1 s. These newly observed collective dynamic responses, due to simultaneous ion channel activation/deactivation by a uniform constant-potential extracellular environment, suggest that perm-selective membrane modules on cell culture chips can facilitate studies of extracellular cardiac cell electrical communication and how ion-channel related pathologies affect cardiac cell synchronization. The future applications of this new technology can lead to better drug screening platforms for cardiotoxicity as well as platforms that can facilitate synchronized maturation of pluripotent stem cells into colonies with high electrical connectivity. 

Liquid biopsy, screening cancer non‐invasively and frequently by detecting and quantifying molecular markers in physiological fluids, would significantly improve cancer survival rate but it remains a distant goal. The key obstacles presented by the highly heterogeneous samples are rapid/high‐yield purification and precise/selective marker capture by their antibody and oligo probes. As irregular expressions of these molecular biomarkers are the key signals, quantifying only those from the cancer cells would greatly enhance the performance of the screening tests. The recent discovery that the biomarkers are carried by nanocarriers, such as exosomes, with cell‐specific membrane proteins suggests that such selection may be possible, although a new suite of fractionation and quantification technologies would need to be developed. Although under‐appreciated, membrane microfluidics has made considerable contributions to resolving these issues. We review the progress made so far, based on ion‐selective, track‐etched, and gel membranes and advanced electrophoretic and nano‐filtration designs, in this perspective and suggest future directions.

 

Current rate of data generation and the need for real‐time data analytics can benefit from new computational approaches where computation proceeds in a massively parallel way while being scalable and energy efficient. Biological systems arising from interaction of living cells can provide such pathways for sustainable computing. Current designs for biocomputing leveraging the information processing units of the cells, such as DNA, gene, or protein circuitries, are inherently slow (hours to days speed) and, therefore, are primarily being considered for archival storage of information. On the contrary, electrically active cells that can synchronize in milliseconds and can be connected as networks to perform massively parallel tasks can transform biocomputing and lead to novel ways of high throughput information processing. Herein, coupled oscillator networks made of living cardiac muscle cells, or bio‐oscillators, is explored as collective computing components for solving computationally hard problems. An empirically validated circuit compatible macromodel is developed for the bio‐oscillators and the fibroblast cells acting as coupling elements, to faithfully reproduce the synchronization dynamics of the network and it is shown that such bio‐oscillator network can be scaled up to hundreds of nodes and be used to solve computationally hard problems faster than traditional heuristics‐based Boolean algorithms.

 

Functionalized nanoparticles (NPs) are the foundation of diverse applications. Especially, in many biosensing applications, concentrating suspended NPs onto a surface without deteriorating their biofunction is usually an inevitable step to improve detection limit, which remains to be a great challenge. In this work, biocompatible deposition of functionalized NPs to optically transparent surfaces is demonstrated using shrinking bubbles. Leveraging the shrinking phase of bubble mitigates the biomolecule degradation problems encountered in traditional photothermal deposition techniques. The deposited NPs are closely packed, and the functional molecules are able to survive the process as verified by their strong fluorescence signals. Using high‐speed videography, it is revealed that the contracting contact line of the shrinking bubble forces the NPs captured by the contact line to a highly concentrated island. Such shrinking surface bubble deposition (SSBD) is low temperature in nature as no heat is added during the process. Using a hairpin DNA‐functionalized gold NP suspension as a model system, SSBD is shown to enable much stronger fluorescence signal compared to the optical‐pressure deposition and the conventional thermal bubble contact line deposition. The demonstrated SSBD technique capable of directly depositing functionalized NPs may significantly simplify biosensor fabrication and thus benefit a wide range of relevant applications.