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In the skeletal muscle tissue, cells are organized following an anisotropic architecture, which is both required during myogenesis when muscle precursor cells fuse to generate myotubes and for its contractile function. To build an in vitro skeletal muscle tissue, it is therefore essential to develop methods to organize cells in an anisotropic fashion, which can be particularly challenging, especially in 3D. In this study, we present a versatile muscle-on-chip system with adjustable collagen hollow tubes that can be seeded with muscle precursor cells. The collagen acts both as a tube-shaped hollow mold and as an extracellular matrix scaffold that can house other cell types for co-culture. We found that the diameter of the channel affects the organization of the muscle cells and that proper myogenesis was obtained at a diameter of 75 μm. In these conditions, muscle precursor cells fused into long myotubes aligned along these collagen channels, resulting in a fascicle-like structure. These myotubes exhibited actin striations and upregulation of multiple myogenic genes, reflecting their maturation. Moreover, we showed that our chip allowed muscle tissue culture and maturation over a month, with the possibility of fibroblast co-culture embedding in the collagen matrix. [ABSTRACT FROM AUTHOR]

Integrin clusters facilitate mechanical force transmission (mechanotransduction) and regulate biochemical signaling during cell adhesion. However, most studies have focused on rigid substrates. On fluid substrates like supported lipid bilayers (SLBs), integrin ligands are mobile, and adhesive complexes are traditionally thought unable to anchor for cell spreading. Here, we demonstrate that cells spread on SLBs coated with Invasin, a high-affinity integrin ligand. Unlike SLBs functionalized with RGD peptides, integrin clusters on Invasin-SLBs grow in size and complexity comparable to those on glass. While actomyosin contraction dominates adhesion maturation on stiff substrates, we find that on fluid SLBs, integrin mechanotransduction and cell spreading rely on dynein pulling forces along microtubules perpendicular to the membranes and microtubules pushing on adhesive complexes, respectively. These forces, potentially present on non-deformable surfaces, are revealed in fluid substrate systems. Supported by a theoretical model, our findings demonstrate a mechanical role for microtubules in integrin clustering., Competing Interests: Competing interests: The authors declare no competing interests., (© 2025. The Author(s).)

The optimization of animal welfare, meat quality, environmental impact, and economic sustainability in alternative poultry farming can be achieved by modulating several productive factors and improving the synergy between the chicken genotype and the outdoor environment. The objective of the study was to characterize 4 slow-growing chicken genotypes reared in free range conditions. Eight hundred chickens (SGs; 25 chickens/replicates/genotype/enrichment) belonging to the following genotypes, Red JA57 (RJ), Naked Neck (NN), Lohmann Dual meat-type (LD), and an Italian crossbreed (Robusta Maculata x Sasso, CB). were utilised and slaughtered at 81 d: The grazing areas were alternatively provided with enrichment constituted by strips of sorghum plants (ENR) or only grass (NO ENR). Productive performance (daily weight gain, daily feed intake, feed conversion ratio, live weight) were recorded weekly. Behaviour observations (walking and grass pecking), carcass and meat quality of breast and drumstick were also assessed in 15 chickens/replicate/genotypes/enrichment. Results demonstrated that both LD and CB showed the highest walking activity, but the different strains were differently capable of using the foraging resources (eating grass). The better productive performance was recorded in RJ followed by NN, CB and LD. In LD and CB, the different walking activities also affected the physico-chemical profiles (lower pHu, WHC, and lipids) of the breast and drumstick. The oxidative status was worse in CB than in the other groups (lower tocols, higher carbonyls), in both meat cuts. Fatty acid profile was also related to the genetic strain: a higher amount of n-3 polyunsaturated fatty acids was recorded both in the breast and drumstick of RJ and NN. The Healthy Fatty Index resulted excellent in all the chicken genotypes. In conclusion, the environment/animal interaction resulted as an important factor affecting the adaptability of genotypes to an extensive rearing system. All four genotypes, to different extents, showed good adaptability and production performance, with the exception of LD and CB, which were too light for the commercial supply chain requirements., Competing Interests: DISCLOSURES The authors declare no conflicts of interest., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

In recent years, the analysis of circulating cell-free DNA (cfDNA) containing tumor-derived DNA has emerged as a noninvasive means for cancer monitoring and personalized medicine. However, the isolation of cfDNA from peripheral blood has remained a challenge due to the low abundance and high fragmentation of these molecules. Here, we present a dynamic Magnetic ExTRactiOn (METRO) protocol using microfluidic fluidized bed technology to isolate circulating cfDNA from raw biological materials such as undiluted serum. This protocol maximizes the surface area for DNA binding within the chip in order to capture short DNA fragments. It uses only a few μL of sample and reagents. The protocol can be automated, and it is fully compatible with sensitive DNA amplification methods such as droplet-based digital PCR (ddPCR)., (© 2024. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)

During invasion, cells breach basement membrane (BM) barriers with actin-rich protrusions. It remains unclear, however, whether actin polymerization applies pushing forces to help break through BM, or whether actin filaments play a passive role as scaffolding for targeting invasive machinery. Here, using the developmental event of anchor cell (AC) invasion in Caenorhabditis elegans , we observe that the AC deforms the BM and underlying tissue just before invasion, exerting forces in the tens of nanonewtons range. Deformation is driven by actin polymerization nucleated by the Arp2/3 complex and its activators, whereas formins and cross-linkers are dispensable. Delays in invasion upon actin regulator loss are not caused by defects in AC polarity, trafficking, or secretion, as appropriate markers are correctly localized in the AC even when actin is reduced and invasion is disrupted. Overall force production emerges from this study as one of the main tools that invading cells use to promote BM disruption in C. elegans ., Competing Interests: The authors declare no conflict of interest., (Copyright © 2018 the Author(s). Published by PNAS.)

Rac1 is a small RhoGTPase switch that orchestrates actin branching in space and time and protrusion/retraction cycles of the lamellipodia at the cell front during mesenchymal migration. Biosensor imaging has revealed a graded concentration of active GTP-loaded Rac1 in protruding regions of the cell. Here, using single-molecule imaging and super-resolution microscopy, we show an additional supramolecular organization of Rac1. We find that Rac1 partitions and is immobilized into nanoclusters of 50-100 molecules each. These nanoclusters assemble because of the interaction of the polybasic tail of Rac1 with the phosphoinositide lipids PIP2 and PIP3. The additional interactions with GEFs and possibly GAPs, downstream effectors, and other partners are responsible for an enrichment of Rac1 nanoclusters in protruding regions of the cell. Our results show that subcellular patterns of Rac1 activity are supported by gradients of signaling nanodomains of heterogeneous molecular composition, which presumably act as discrete signaling platforms., (Copyright © 2017 The Author(s). Published by Elsevier Inc. All rights reserved.)