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On-Chip Quantitative Measurement of Mechanical Stresses During Cell Migration with Emulsion Droplets
D. Molino, S. Quignard, C. Gruget, F. Pincet, Y. Chen, M. Piel & J. Fattaccioli
Scientific Reports - 6 29113 - DOI: 10.1038/srep29113 - 2019
The ability of immune cells to migrate within narrow and crowded spaces is a critical feature involved in various physiological processes from immune response to metastasis. Several in-vitro techniques have been developed so far to study the behaviour of migrating cells, the most recent being based on the fabrication of microchannels within which cells move. To address the question of the mechanical stress a cell is able to produce during the encounter of an obstacle while migrating, we developed a hybrid microchip made of parallel PDMS channels in which oil droplets are sparsely distributed and serve as deformable obstacles. We thus show that cells strongly deform droplets while passing them. Then, we show that the microdevice can be used to study the influence of drugs on migration at the population level. Finally, we describe a quantitative analysis method of the droplet deformation that allows measuring in real-time the mechanical stress exerted by a single cell. The method presented herein thus constitutes a powerful analytical tool for cell migration studies under confinement.
Nano-on-Micro Fibrous Extracellular Matrices for Scalable Expansion of Human Es/Ips Cells
L. Liu, K.-i. Kamei, M. Yoshioka, M. Nakajima, J. Li, N. Fujimoto, S. Terada, Y. Tokunaga, Y. Koyama, H. Sato, K. Hasegawa, N. Nakatsuji and Y. Chen
Biomaterials - 124 47-54 - DOI: 10.1016/j.biomaterials.2017.01.039 - 2019
Human pluripotent stem cells (hPSCs) hold great potential for industrial and clinical applications. Clinical-grade scaffolds and high-quality hPSCs are required for cell expansion as well as easy handling and manipulation of the products. Current hPSC culture methods do not fulfill these requirements because of a lack of proper extracellular matrices (ECMs) and cell culture wares. We developed a layered nano-on-micro fibrous cellular matrix mimicking ECM, named "fiber-on-fiber (FF)" matrix, which enables easy handling and manipulation of cultured cells. While non-woven sheets of cellulose and polyglycolic acid were used as a microfiber layer facilitating mechanical stability, electrospun gelatin nanofibers were crosslinked on the microfiber layer, generating a mesh structure with connected nanofibers facilitating cell adhesion and growth. Our results showed that the FF matrix supports effective hPSC culture with maintenance of their pluripotency and normal chromosomes over two months, as well as effective scaled-up expansion, with fold increases of 54.1 ± 15.6 and 40.4 ± 8.4 in cell number per week for H1 human embryonic stem cells and 253G1 human induced pluripotent stem cells, respectively. This simple approach to mimick the ECM may have important implications after further optimization to generate lineage-specific products.
RAB-35 and ARF-6 GTPases Mediate Engulfment and Clearance Following Linker Cell-Type Death
Kutscher LM, Keil W, Shaham S
Dev Cell - 47(2) 222-238 - doi: 10.1016/j.devcel.2018.08.015. - 2018
Clearance of dying cells is essential for development and homeostasis. Conserved genes mediate apoptotic cell removal, but whether these genes control non-apoptotic cell removal is a major open question. Linker cell-type death (LCD) is a prevalent non-apoptotic developmental cell death process with features conserved from C. elegans to vertebrates. Using microfluidics-based long-term in vivo imaging, we show that unlike apoptotic cells, the C. elegans linker cell, which dies by LCD, is competitively phagocytosed by two neighboring cells, resulting in cell splitting. Subsequent cell elimination does not require apoptotic engulfment genes. Rather, we find that RAB-35 GTPase is a key coordinator of competitive phagocytosis onset and cell degradation. RAB-35 binds CNT-1, an ARF-6 GTPase activating protein, and removes ARF-6, a degradation inhibitor, from phagosome membranes. This facilitates phosphatidylinositol-4,5-bisphosphate removal from phagosome membranes, promoting phagolysosome maturation. Our studies suggest that RAB-35 and ARF-6 drive a conserved program eliminating cells dying by LCD.
Selection Dynamics in Transient Compartmentalization.
Blokhuis A, Lacoste D, Nghe P, Peliti L
Phys. Rev. Lett. - 158101 120(15): - doi: 10.1371/journal.pcbi.1004972 - 2018
Transient compartments have been recently shown to be able to maintain functional replicators in the context of prebiotic studies. Here, we show that a broad class of selection dynamics is able to achieve this goal. We identify two key parameters, the relative amplification of nonactive replicators (parasites) and the size of compartments. These parameters account for competition and diversity, and the results are relevant to similar multilevel selection problems, such as those found in virus-host ecology and trait group selection.
Coupled catabolism and anabolism in autocatalytic RNA sets.
Arsène S, Ameta S, Lehman N, Griffiths AD, Nghe P.
Nucleic Acids Res. - 46(18) 9660-9666 - doi: 10.1093/nar/gky598. - 2018
The ability to process molecules available in the environment into useable building blocks characterizes catabolism in contemporary cells and was probably critical for the initiation of life. Here we show that a catabolic process in collectively autocatalytic sets of RNAs allows diversified substrates to be assimilated. We modify fragments of the Azoarcus group I intron and find that the system is able to restore the original native fragments by a multi-step reaction pathway. This allows in turn the formation of catalysts by an anabolic process, eventually leading to the accumulation of ribozymes. These results demonstrate that rudimentary self-reproducing RNA systems based on recombination possess an inherent capacity to assimilate an expanded repertoire of chemical resources and suggest that coupled catabolism and anabolism could have arisen at a very early stage in primordial living systems.
Sign epistasis caused by hierarchy within signalling cascades
Nghe P, Kogenaru M, Tans SJ
Nat Commun - 1451 9660-9666 - 10.1038/s41467-018-03644-8. - 2018
The ability to process molecules available in the environment into useable building blocks characterizes catabolism in contemporary cells and was probably critical for the initiation of life. Here we show that a catabolic process in collectively autocatalytic sets of RNAs allows diversified substrates to be assimilated. We modify fragments of the Azoarcus group I intron and find that the system is able to restore the original native fragments by a multi-step reaction pathway. This allows in turn the formation of catalysts by an anabolic process, eventually leading to the accumulation of ribozymes. These results demonstrate that rudimentary self-reproducing RNA systems based on recombination possess an inherent capacity to assimilate an expanded repertoire of chemical resources and suggest that coupled catabolism and anabolism could have arisen at a very early stage in primordial living systems.
Selection Dynamics in Transient Compartmentalization
doi.org/10.1103/PhysRevLett.120.158101
Phys. Rev. Lett. - 120 158101 - doi.org/10.1103/PhysRevLett.120.158101 - 2018
Transient compartments have been recently shown to be able to maintain functional replicators in the context of prebiotic studies. Here, we show that a broad class of selection dynamics is able to achieve this goal. We identify two key parameters, the relative amplification of nonactive replicators (parasites) and the size of compartments. These parameters account for competition and diversity, and the results are relevant to similar multilevel selection problems, such as those found in virus-host ecology and trait group selection.
Electrostrictive polymer composites based on liquid crystalline graphene for mechanical energy harvesting
Jinkai Yuan, Wilfrid Neri, Cécile Zakri, Philippe Poulin, Annie Colin
Multifunctional Materials - 1-4 - - 2018
High electromechanical coupling is critical to perform effective conversion between mechanical and electrical energy for various applications of electrostrictive polymers. Herein, a giant electrostriction effect is reported in liquid crystalline graphene doped dielectric elastomers. The materials are formulated by an original phase transfer method which allows the solubilization of graphenic monolayers in non-polar solvents. Dielectric spectroscopy is combined with tensile test devices to measure the true electrostriction coefficients with differentiating the Maxwell stress effect. Because of their unique liquid crystal structure, the resultant composites show a giant relative permittivity and ultralarge electrostriction coefficient. This work offers a promising pathway to design novel high performance electrostrictive polymer composites as well as to provide insights into mechanisms of true electrostriction in electrically …
Microporous electrostrictive materials for vibrational energy harvesting
Mickaël Pruvost, Wilbert J Smit, Cécile Monteux, Philippe Poulin, Annie Colin
Multifunctional Materials - 1 015004 - - 2018
We present electrostrictive materials with excellent properties for vibrational energy harvesting applications. The developed materials consist of a porous carbon black composite, which is processed using water-in-oil emulsions. In combination with an insulating layer, the investigated structures exhibit a high effective relative dielectric permittivity (up to 182 at 100 Hz) with very low effective conductivity (down to 2.53 10− 8 S m− 1). They can generate electrical energy in response to mechanical vibrations with a power density of 0.38 W m− 3 under an applied bias electric field of 32 V. They display figures or merit for energy harvesting applications well above reference polymer materials in the field, including fluorinated co-and ter-polymers synthetized by heavy chemical processes. The production process of the present materials is based on non hazardous and low-cost chemicals. The soft dielectric materials are …
Shear Rheology Control of Wrinkles and Patterns in Graphene Oxide Films
Franco Tardani, Wilfrid Neri, Cécile Zakri, Hamid Kellay, Annie Colin, Philippe Poulin
Langmuir - 9(34) 2996-3002 - - 2018
Drying graphene oxide (GO) films are subject to extensive wrinkling, which largely affects their final properties. Wrinkles were shown to be suitable in biotechnological applications; however, they negatively affect the electronic properties of the films. Here, we report on wrinkle tuning and patterning of GO films under stress-controlled conditions during drying. GO flakes assemble at an air–solvent interface; the assembly forms a skin at the surface and may bend due to volume shrinkage while drying. We applied a modification of evaporative lithography to spatially define the evaporative stress field. Wrinkle alignment is achieved over cm2 areas. The wavelength (i.e., wrinkle spacing) is controlled in the μm range by the film thickness and GO concentration. Furthermore, we propose the use of nanoparticles to control capillary forces to suppress wrinkling. An example of a controlled pattern is given to elucidate the …

400 publications.