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Uniflex culture plates

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The UniFlex culture plates are a versatile laboratory equipment designed for cell culture applications. They feature a uniform, flexible substrate that allows for the application of mechanical strain to cultured cells. The plates are intended to facilitate the study of cellular responses to mechanical stimuli.

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Lab products found in correlation

Fx 5000 tension system, by Flexcell (4 mentions) Fx 6000, by Flexcell (3 mentions) Pkh26 red fluorescent cell linker, by Merck Group (1 mentions) Hepes, by MP Biomedicals (1 mentions) Boyden chamber, by Corning (1 mentions) Biocoat matrigel invasion chamber, by Corning (1 mentions) Mtesr e8 media, by STEMCELL (1 mentions) Vitronectin xf, by STEMCELL (1 mentions)

7 protocols using uniflex culture plates

1

Exosome Release Quantification by Biomechanical Stress

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The rate of release of exosomes is influenced by biomechanical forces acting on the tumor cells, and to quantify this, an experimental setup depicted in Figure 1 was used. The murine TNBC cell line 4T1.2 (an aggressive clone derived from 4T1) was obtained from Dr. Robin L. Anderson's laboratory (Peter McCallum Cancer Center, Australia). 4T1.2 cells were cultured in Dulbecco's Modified Eagle Medium (DMEM) supplemented with 10% FBS and 10 mM HEPES (MP Biomedicals, Santa Ana, CA). Prior to the exposure to tensile strain, the 4T1.2 cells were stained with the lypophilic dye PKH26 Red Fluorescent Cell Linker (Sigma, St. Louis, MO), per the manufacturer's instructions. 2.5 × 105 4T1.2 were seeded on collagen coated 6 well UniFlex culture plates (Flexcell International Corporation, Burlington, NC) and cultured to confluence. Once confluent, the media was changed to exosome depleted growth media and the plates were subjected to 10% uniaxial oscillatory strain at 0.3 Hz for 48 h, 10% constant strain for 48 h, or no strain for 48 h using a FlexCell FX-6000 or FX-5000 Tension System.
Koomullil R., Tehrani B., Goliwas K., Wang Y., Ponnazhagan S., Berry J, & Deshane J. (2021). Computational Simulation of Exosome Transport in Tumor Microenvironment. Frontiers in Medicine, 8, 643793.
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2

Boyden Chamber Cell Migration Assay

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A commercially available Boyden chamber (Corning®, NY, USA) was used to detect cell migration. The PET Membrane was 8.0 μm. After undergoing mechanical strain for 3 days, cancer cells were trypsinized form the Flexcell Uniflex® Culture Plates, centrifuged, and resuspended in serum free RPMI and counted. An equal number of cells (5 × 104 cells/well) were loaded onto the upper chamber. Medium enriched with 10% FBS was added in the lower chamber as a chemoattractant. After incubation (37 °C, 5% CO2) for 24 h, non-migrated cells remained on the upper surface of the membrane and were removed; cells that migrated through the membrane were fixed with formalin and stained with Crystal violet (Fischer chemicals). Cells that had migrated were counted under a light microscope.
Martinez A., Buckley M., Scalise C.B., Katre A.A., Dholakia J.J., Crossman D., Birrer M.J., Berry J.L, & Arend R.C. (2021). Understanding the effect of mechanical forces on ovarian cancer progression. Gynecologic oncology, 162(1), 154-162.
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3

Matrigel Invasion Assay for Cancer Cells

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A commercially available BioCoat™ Matrigel® Invasion Chamber (Corning®, NY, USA) was used to measure cell invasion. The PET Membrane was 8.0 μm and coated with Matrigel®. After undergoing mechanical strain for 3 days, cancer cells were trypsinized form the Flexcell Uniflex® Culture Plates, centrifuged, and resuspended in serum free RPMI and counted. An equal number of cells (5 × 104 cells/well) were loaded onto the upper chamber of the invasion assay. Medium enriched with 10% FBS was added in the lower chamber as a chemoattractant. After incubation (37 °C, 5% CO2) for 24 h, non-invaded cells remained on the upper surface of the membrane and were removed; cells that invaded through the membrane were fixed with formalin and stained with Crystal violet (Fischer chemicals)Cells that had invaded were counted under a light microscope.
Martinez A., Buckley M., Scalise C.B., Katre A.A., Dholakia J.J., Crossman D., Birrer M.J., Berry J.L, & Arend R.C. (2021). Understanding the effect of mechanical forces on ovarian cancer progression. Gynecologic oncology, 162(1), 154-162.
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4

Mechanical Strain Induces Cancer Cell Response

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2.5 × 105 BCa cells (MCF-7-GFP/LUC, MDA-MB-231-GFP/LUC or 4T1.2) were seeded on collagen coated 6 well UniFlex culture plates (Flexcell International Corporation, Burlington, NC) and cultured to confluence in the growth medium appropriate for each cell line. Using a FlexCell FX-6000 or FX-5000 Tension System, plates were subjected to 10% uniaxial oscillatory strain at 0.3 Hz for 48 hours, 10% constant strain for 48 hours, or no strain for 48 hours with medium changed immediately prior to induction of strain.
Wang Y., Goliwas K.F., Severino P.E., Hough K., Van Vessem D., Wang H., Tousif S., Koomullil R.P., Frost A.R., Ponnazhagen S., Berry J.L, & Deshane J.S. (2020). Mechanical Strain Induces Phenotypic Changes in Breast Cancer Cells and Promotes Immunosuppression in the Tumor Microenvironment. Laboratory investigation; a journal of technical methods and pathology, 100(12), 1503-1516.
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5

Mechanical Strain Induces Cancer Cell Response

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2.5 × 105 BCa cells (MCF-7-GFP/LUC, MDA-MB-231-GFP/LUC or 4T1.2) were seeded on collagen coated 6 well UniFlex culture plates (Flexcell International Corporation, Burlington, NC) and cultured to confluence in the growth medium appropriate for each cell line. Using a FlexCell FX-6000 or FX-5000 Tension System, plates were subjected to 10% uniaxial oscillatory strain at 0.3 Hz for 48 hours, 10% constant strain for 48 hours, or no strain for 48 hours with medium changed immediately prior to induction of strain.
Wang Y., Goliwas K.F., Severino P.E., Hough K., Van Vessem D., Wang H., Tousif S., Koomullil R.P., Frost A.R., Ponnazhagen S., Berry J.L, & Deshane J.S. (2020). Mechanical Strain Induces Phenotypic Changes in Breast Cancer Cells and Promotes Immunosuppression in the Tumor Microenvironment. Laboratory investigation; a journal of technical methods and pathology, 100(12), 1503-1516.
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6

Stretch-induced iPSC-CM Maturation

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Flexcell(R) Tissue Train(R) Culture System was set up in a regular incubator according to manufacturer’s instructions. Commercially available laminin coated UniFlex® culture plates (Flexcell) with PDMS membrane bottom were prepared. Human iPSC-CMs, following purification by lactate conditioned medium, were seeded in 6-well UniFlex® culture plates according to manufacture’s instructions. When iPSC-CMs formed a confluent monolayer culture, unidirectional stretch, set at 5% elongation, was carried out for 7 days at 0.5 Hz. Following the stretching period, cells were cultured in regular static condition until their detachment and seeding in printed constructs.
Ma X., Dewan S., Liu J., Tang M., Miller K.L., Yu C., Lawrence N., McCulloch A.D, & Chen S. (2018). 3D Printed Micro-Scale Force Gauge Arrays to Improve Human Cardiac Tissue Maturation and Enable High Throughput Drug Testing. Acta biomaterialia, 95, 319-327.
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7

CRISPR-Corrected iPSCs Differentiated into Stretched SMCs

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Dermal fibroblasts from a patient with MFS (Coriell Institute, GM21943) were reprogrammed into human induced pluripotent stem cells (iPSCs), and the mutation was corrected by CRISPR-Cas9 to generate an isogenic control.26 (link) Control and Marfan iPSCs were grown on Vitronectin XF (STEMCELL Technologies) and maintained in mTeSR E8 media (STEMCELL Technologies). They were then differentiated into neural crest SMCs using the methods described previously.27 (link),28 (link) After differentiation, the resulting SMCs were maintained in DMEM supplemented with 10% fetal bovine serum for at least 2 weeks before seeding onto UniFlex Culture Plates (FlexCell International Corporation). After 4 days, the SMCs were stretched for 48 hours using a Cyclic Stress Unit (FX5000 Tension System, FlexCell International Corporation), using a cyclic sine wave (60 cycles/min) and 10% elongation.
Yin (殷晓科) X., Wanga S., Fellows A.L., Barallobre-Barreiro J., Lu R., Davaapil H., Franken R., Fava M., Baig F., Skroblin P., Xing Q., Koolbergen D.R., Groenink M., Zwinderman A.H., Balm R., de Vries C.J., Mulder B.J., Viner R., Jahangiri M., Reinhardt D.P., Sinha S., de Waard V, & Mayr M. (2019). Glycoproteomic Analysis of the Aortic Extracellular Matrix in Marfan Patients. Arteriosclerosis, Thrombosis, and Vascular Biology, 39(9), 1859-1873.
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