Fx 3000 strain unit

Manufactured by Flexcell

Sourced in United States

The FX-3000 Flexcell Strain Unit is a laboratory equipment designed to apply controlled mechanical strain to cell cultures. It provides a consistent and reproducible method for stretching or compressing cells in vitro. The device utilizes a vacuum-based system to deform flexible-bottomed culture plates or dishes, allowing for the application of adjustable strain patterns to the cultured cells.

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Lab products found in correlation

Collagen type 1, by Flexcell (2 mentions) Bioflex plate, by Flexcell (2 mentions) No bf 3001c, by Flexcell (2 mentions) Interferon gamma, by Thermo Fisher Scientific (1 mentions) Bioflex, by Flexcell (1 mentions) Bioflex culture plate, by Flexcell (1 mentions) Fetal bovine serum (fbs), by Biowest (1 mentions) Mouse osteogenesis culture kit, by Cosmo Bio (1 mentions)

Spelling variants of this product

Flexercell Strain Unit FX-3000 (6 citations) (FX-3000; (2 citations)

8 protocols using fx 3000 strain unit

Mechanical Stretch-Induced Cytokine Secretion

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EC were plated onto six-well silicone elastomer Bioflex plates coated with type I collagen (FlexCell International, Hillsborough, NC) and grown to 75–80% prior to transfection as described. Mechanical stretch was performed via the Flexcell Strain Unit (FX-3000; FlexCell International) placed in a 5% CO₂ incubator at 37 °C and 95% humidity. The device uses a controlled vacuum to induce CS with 18% elongation at a frequency of 30 cycles per minute (0.5 Hz) for 6 h. The media was then collected and briefly centrifuged to measure cytokine levels with Bio-Rad Bio-Plex ELISA kits (Hercules, CA) according to the manufacturer’s instructions.

Chen W., Epshtein Y., Ni X., Dull R.O., Cress A.E., Garcia J.G, & Jacobson J.R. (2015). Role of Integrin β4 in Lung Endothelial Cell Inflammatory Responses to Mechanical Stress. Scientific Reports, 5, 16529.

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Lung Epithelial Cell Stretch Assay

Cited 1 time

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Immortalized mouse lung epithelial cells (MLE-15) were the generous gift of Jeffrey Whitsett, M.D. (Professor of Pediatrics, University of Cincinnati College of Medicine, Cincinnati, OH) and were cultured as described previously.^{30 (link)} For cyclic stretch experiments, cells were plated on Pronectin coated Flexcell plates (Flexcell, Hillsborough, NC) and grown to confluence. Cells were then treated with a combination of 0.5 μg/ml Escherichia coli lipopolysaccharide (L2762, Sigma-Aldrich) and 100 units/ml of interferon gamma (315-05, Peprotech, Rocky Hill, NJ) or PBS control for 4 hours (Fig 1B). Cells were subsequently exposed to isoflurane (or control gas containing 23% oxygen, 5% CO₂, balance nitrogen) in an airtight chamber for 2 hours as described previously.^{31 (link)} Isoflurane, oxygen, and carbon dioxide concentrations were measured every 30 minutes as described previously.^{31 (link)} After isoflurane exposure, cell were incubated overnight prior to biaxial cyclic stretch (10% stretch, 0.5 Hz) delivered via a Flexcell Strain Unit FX-3000 (Flexcell) for 2 hours prior to isolating RNA or protein. Control cells were placed in the stretching device, but not subjected to stretch.

Englert J.A., Macias A.A., Amador-Munoz D., Vera M.P., Isabelle C., Guan J., Magaoay B., Velandia M.S., Coronata A., Lee A., Fredenburgh L.E., Culley D.J., Crosby G, & Baron R.M. (2015). Isoflurane ameliorates acute lung injury by preserving epithelial tight junction integrity. Anesthesiology, 123(2), 377-388.

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Endothelial Cell Response to LPS and Cyclic Stretch

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For the LPS challenge, ECs were cultured in EGM-2 and challenged with 100 ng/ml of LPS for various time points as indicated. The resulting media were collected and briefly centrifuged using ELISA assay (Biolegend, San Diego, CA, United States) according to the manufacturer’s instructions. For CS experiments, ECs were plated onto six-well silicone elastomer Bioflex plates coated with type I collagen (FlexCell International, Hillsborough, NC, United States) and grown to 75–80% prior to transfection as described. Mechanical stretch was performed via the Flexcell Strain Unit (FX-3000; FlexCell International) placed in a 5% of CO₂ incubator at 37°C and 95% of humidity. The device uses a controlled vacuum to induce CS with 18% of elongation at a frequency of 30 cycles per minute (0.5 Hz) for 4 h. The media were then collected and briefly centrifuged prior to the ELISA assay.

Chen W., Gard J.M., Epshtein Y., Camp S.M., Garcia J.G., Jacobson J.R, & Cress A.E. (2022). Integrin Beta 4E Promotes Endothelial Phenotypic Changes and Attenuates Lung Endothelial Cell Inflammatory Responses. Frontiers in Physiology, 13, 769325.

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Mechanical Stretch Regulates LPAR and LPP Expression

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Primary cultures of HTM cells were plated onto BioFlex six-well plates precoated with collagen type I (Flexcell International Corp., Burlington, NC, USA). As the cells reached confluence, cultured medium was switched to serum-free, phenol-free DMEM, and cells were subjected to cyclic mechanical stretch (15% stretching, 1 cycle/sec) for 24 hours, using the computer-controlled vacuum-operated FX-3000 Flexcell Strain Unit (Flexcell, Hillsborough, NC, USA), as we previously described.^{10 (link)} Control cells were cultured under similar conditions with no mechanical force applied. RNA was extracted from the cells and analyzed by RT-quantitative (q)PCR to monitor changes in level of expression of LPARs, LPPs, and ATX.

Ho L.T., Skiba N., Ullmer C, & Rao P.V. (2018). Lysophosphatidic Acid Induces ECM Production via Activation of the Mechanosensitive YAP/TAZ Transcriptional Pathway in Trabecular Meshwork Cells. Investigative Ophthalmology & Visual Science, 59(5), 1969-1984.

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Mechanical Stretching of Osteoblasts

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The experimental osteoblasts were induced from the mouse bone marrow osteogenic stromal cells (Cosmo Bio Co., LTD, Tokyo, Japan) with a mouse osteogenesis culture kit (Cosmo Bio) according to the supplier recommendations. Osteoblasts were maintained in Minimum Essential Medium Eagle, Alpha Modification (α-MEM, Sigma-Aldrich Co. LLC., St. Louis, MO) with 10% fetal bovine serum (Biowest, Nuaillé, France). The cells (10,000 cells/well) were seeded on collagen I-coated elastic silicon membranes of 6-well BioFlex Culture Plates (25-mm diameter, 9.6-cm² well; Flexcell International Corp, Burlington, NC) in the mineralization medium containing 100 nM dexamethasone, 50 μg/ml, ascorbic acid, and 10 mM β-glycerophosphate for the osteogenic differentiation to test the mineralization. When cells reached about 80% confluence, the cells were subjected to vertical stretching on silicon membrane (5% elongation at 2 cycles/min) using a FX-3000 Flexcell Strain Unit (Flexcell) for five days.

Takenawa T., Kanai T., Kitamura T., Yoshimura Y., Sawa Y, & Iida J. (2018). Expression and Dynamics of Podoplanin in Cultured Osteoblasts with Mechanostress and Mineralization Stimulus. Acta Histochemica et Cytochemica, 51(1), 41-52.

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Cyclic Stretch and RBC Supernatant Effects

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A549 cells were incubated with supernatant from fresh (day 4–5) and old (day 41–42) RBC products and subjected to 25% stretch at a frequency of 12 cycles/min (0.2 Hz) using a computer‐driven FX‐3000 Flexcell strain unit (Flexcell International) for 24 h at 37°C in the presence of 5% CO₂. These parameters reflect mechanical ventilation with high tidal volumes and are based on previously published methods [9] and pilot studies. The device creates a vacuum below the flexible membrane which deforms the cells grown on top of the membrane to mimic the stress on pulmonary cells during mechanical ventilation. Control cells were either cultured on the flexible membranes, but not subjected to stretch; or subjected to stretch, but not incubated with supernatant of RBC products. Supernatant of the stretched and non‐stretched cells was collected after 24 h of (cyclic stretch) exposure.

Wirtz M.R., Almizraq R.J., Weber N.C., Norris P.J., Pandey S., Spinella P.C., Muszynski J.A., P. Acker J, & Juffermans N.P. (2020). Red‐blood‐cell manufacturing methods and storage solutions differentially induce pulmonary cell activation. Vox Sanguinis, 115(5), 395-404.

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Mechanically Induced GDF-15 Expression

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Primary cultures of human TM cells were plated on collagen Type 1 BioFlex culture plates with a flexible silicone bottom (C. No. BF-3001C, Flexcell International Corporation, Burlington, NC, USA). As the cells reached confluence, culture medium was switched to serum-free, phenol-free DMEM, and cells were subjected to cyclic mechanical stretch (20% stretching, one cycle per second) for 48 hours, using the computer-controlled, vacuum-operated FX-3000 Flexcell Strain Unit (Flexcell, Hillsborough, NC, USA) as we described previously.⁹ Control cells were cultured under similar conditions with no mechanical force applied. Conditioned media were collected from control and stretched cells to analyze GDF-15 protein levels, and RNA was extracted to monitor changes in GDF-15 expression by RT-qPCR analysis.

Muralidharan A.R., Maddala R., Skiba N.P, & Rao P.V. (2016). Growth Differentiation Factor-15–Induced Contractile Activity and Extracellular Matrix Production in Human Trabecular Meshwork Cells. Investigative Ophthalmology & Visual Science, 57(15), 6482-6495.

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Mechanical Strain Induces Secretion

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Primary cultures of human TM cells were plated on collagen Type 1 BioFlex culture plates with flexible silicone bottom (C. No. BF-3001C, Flexcell® International Corporation, Burlington, NC). As the cells reached confluence, culture medium was switched to serum-free phenol free DMEM, and cells were subjected to cyclic mechanical stretch (20% stretching, one cycle per second) for 48 h, using the computer-controlled, vacuum-operated FX-3000 Flexcell Strain Unit (Flexcell, Hillsborough, NC). Control cells were cultured under similar conditions with no mechanical force applied. Conditioned media were collected from both control and stretched cells to analyze the VLK protein levels.

Maddala R., Skiba N.P, & Rao P.V. (2017). Vertebrate Lonesome Kinase Regulated Extracellular Matrix Protein Phosphorylation, Cell Shape and Adhesion in Trabecular Meshwork Cells. Journal of cellular physiology, 232(9), 2447-2460.

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