TY - JOUR A1 - Mai, Tobias A1 - Wolski, Karol A1 - Puciul-Malinowska, Agnieszka A1 - Kopyshev, Alexey A1 - Gräf, Ralph A1 - Bruns, Michael A1 - Zapotoczny, Szczepan A1 - Taubert, Andreas T1 - Anionic polymer brushes for biomimetic calcium phosphate mineralization BT - A surface with application potential in biomaterials JF - Polymers N2 - This article describes the synthesis of anionic polymer brushes and their mineralization with calcium phosphate. The brushes are based on poly(3-sulfopropyl methacrylate potassium salt) providing a highly charged polymer brush surface. Homogeneous brushes with reproducible thicknesses are obtained via surface-initiated atom transfer radical polymerization. Mineralization with doubly concentrated simulated body fluid yields polymer/inorganic hybrid films containing AB-Type carbonated hydroxyapatite (CHAP), a material resembling the inorganic component of bone. Moreover, growth experiments using Dictyostelium discoideum amoebae demonstrate that the mineral-free and the mineral-containing polymer brushes have a good biocompatibility suggesting their use as biocompatible surfaces in implantology or related fields. KW - polymer brushes KW - calcium phosphate KW - hydroxyapatite KW - carbonated apatite KW - bone mimic KW - biocompatibility KW - Dictyostelium discoideum Y1 - 2018 U6 - https://doi.org/10.3390/polym10101165 SN - 2073-4360 VL - 10 IS - 10 PB - MDPI CY - Basel ER - 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 - 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 - TY - JOUR A1 - Barbosa Pfannes, Eva Katharina A1 - Theves, Matthias A1 - Wegner, Christian A1 - Beta, Carsten T1 - Impact of the carbazole derivative wiskostatin on mechanical stability and dynamics of motile cells JF - JOURNAL OF MUSCLE RESEARCH AND CELL MOTILITY N2 - Many essential functions in eukaryotic cells like phagocytosis, division, and motility rely on the dynamical properties of the actin cytoskeleton. A central player in the actin system is the Arp2/3 complex. Its activity is controlled by members of the WASP (Wiskott-Aldrich syndrome protein) family. In this work, we investigated the effect of the carbazole derivative wiskostatin, a recently identified N-WASP inhibitor, on actin-driven processes in motile cells of the social ameba . Drug-treated cells exhibited an altered morphology and strongly reduced pseudopod formation. However, TIRF microscopy images revealed that the overall cortical network structure remained intact. We probed the mechanical stability of wiskostatin-treated cells using a microfluidic device. While the total amount of F-actin in the cells remained constant, their stiffness was strongly reduced. Furthermore, wiskostatin treatment enhanced the resistance to fluid shear stress, while spontaneous motility as well as chemotactic motion in gradients of cAMP were reduced. Our results suggest that wiskostatin affects the mechanical integrity of the actin cortex so that its rigidity is reduced and actin-driven force generation is impaired. KW - Actin dynamics KW - Wiskostatin KW - Dictyostelium discoideum Y1 - 2012 U6 - https://doi.org/10.1007/s10974-012-9287-8 SN - 0142-4319 VL - 33 IS - 2 SP - 95 EP - 106 PB - SPRINGER CY - DORDRECHT 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 -