TY - JOUR A1 - Nagel, Oliver A1 - Frey, Manuel A1 - Gerhardt, Matthias A1 - Beta, Carsten T1 - Harnessing Motile Amoeboid Cells as Trucks for Microtransport and -Assembly JF - Advanced science N2 - Cell-driven microtransport is one of the most prominent applications in the emerging field of biohybrid systems. While bacterial cells have been successfully employed to drive the swimming motion of micrometer-sized cargo particles, the transport capacities of motile adherent cells remain largely unexplored. Here, it is demonstrated that motile amoeboid cells can act as efficient and versatile trucks to transport microcargo. When incubated together with microparticles, cells of the social amoeba Dictyostelium discoideum readily pick up and move the cargo particles. Relying on the unspecific adhesive properties of the amoeba, a wide range of different cargo materials can be used. The cell-driven transport can be directionally guided based on the chemotactic responses of amoeba to chemoattractant gradients. On the one hand, the cargo can be assembled into clusters in a self-organized fashion, relying on the developmentally induced chemotactic aggregation of cells. On the other hand, chemoattractant gradients can be externally imposed to guide the cellular microtrucks to a desired location. Finally, larger cargo particles of different shapes that exceed the size of a single cell by more than an order of magnitude, can also be transported by the collective effort of large numbers of motile cells. KW - biohybrid microsystems KW - chemotaxis KW - Dictyostelium discoideum KW - microtransport and -assembly Y1 - 2018 U6 - https://doi.org/10.1002/advs.201801242 SN - 2198-3844 VL - 6 IS - 3 PB - Wiley CY - Hoboken ER - TY - JOUR A1 - Westendorf, Christian A1 - Negrete, Jose A1 - Bae, Albert J. A1 - Sandmann, Rabea A1 - Bodenschatz, Eberhard A1 - Beta, Carsten T1 - Actin cytoskeleton of chemotactic amoebae operates close to the onset of oscillations JF - Proceedings of the National Academy of Sciences of the United States of America N2 - The rapid reorganization of the actin cytoskeleton in response to external stimuli is an essential property of many motile eukaryotic cells. Here, we report evidence that the actin machinery of chemotactic Dictyostelium cells operates close to an oscillatory instability. When averaging the actin response of many cells to a short pulse of the chemoattractant cAMP, we observed a transient accumulation of cortical actin reminiscent of a damped oscillation. At the single-cell level, however, the response dynamics ranged from short, strongly damped responses to slowly decaying, weakly damped oscillations. Furthermore, in a small subpopulation, we observed self-sustained oscillations in the cortical F-actin concentration. To substantiate that an oscillatory mechanism governs the actin dynamics in these cells, we systematically exposed a large number of cells to periodic pulse trains of different frequencies. Our results indicate a resonance peak at a stimulation period of around 20 s. We propose a delayed feedback model that explains our experimental findings based on a time-delay in the regulatory network of the actin system. To test the model, we performed stimulation experiments with cells that express GFP-tagged fusion proteins of Coronin and actin-interacting protein 1, as well as knockout mutants that lack Coronin and actin-interacting protein 1. These actin-binding proteins enhance the disassembly of actin filaments and thus allow us to estimate the delay time in the regulatory feedback loop. Based on this independent estimate, our model predicts an intrinsic period of 20 s, which agrees with the resonance observed in our periodic stimulation experiments. KW - Dictyostelium discoideum KW - microfluidics KW - caged cAMP KW - delay-differential equation Y1 - 2013 U6 - https://doi.org/10.1073/pnas.1216629110 SN - 0027-8424 VL - 110 IS - 10 SP - 3853 EP - 3858 PB - National Acad. of Sciences CY - Washington ER - TY - JOUR A1 - Beta, Carsten A1 - Bodenschatz, Eberhard T1 - Microfluidic tools for quantitative studies of eukaryotic chemotaxis JF - European journal of cell biology N2 - Over the past decade, microfluidic techniques have been established as a versatile platform to perform live cell experiments under well-controlled conditions. To investigate the directional responses of cells, stable concentration profiles of chemotactic factors can be generated in microfluidic gradient mixers that provide a high degree of spatial control. However, the times for built-up and switching of gradient profiles are in general too slow to resolve the intracellular protein translocation events of directional sensing of eukaryotes. Here, we review an example of a conventional microfluidic gradient mixer as well as the novel flow photolysis technique that achieves an increased temporal resolution by combining the photo-activation of caged compounds with the advantages of microfluidic chambers. KW - Eukaryotic chemotaxis KW - Dictyostelium discoideum KW - Microfluidics KW - Caged compounds KW - Numerical simulations Y1 - 2011 U6 - https://doi.org/10.1016/j.ejcb.2011.05.006 SN - 0171-9335 VL - 90 IS - 10 SP - 811 EP - 816 PB - Elsevier CY - Jena ER -