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Transwell plate

Manufactured by Thermo Fisher Scientific
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The Transwell plate is a cell culture insert system designed for in vitro studies of cell migration, invasion, and permeability. The system consists of a upper and lower chamber separated by a porous membrane, allowing the movement of cells and molecules between the chambers to be monitored.

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Lab products found in correlation

Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions) Ccl20 chemokine, by R&D Systems (2 mentions) Vybrant fluorescent dye, by Thermo Fisher Scientific (2 mentions) Metavue software, by Molecular Devices (2 mentions) Bca protein assay kit, by Thermo Fisher Scientific (2 mentions) Casy tt cell counter, by Roche (1 mentions) Bovine serum albumin (bsa), by R&D Systems (1 mentions) Fibronectin, by Merck Group (1 mentions) Tryple, by Thermo Fisher Scientific (1 mentions) Corticosterone, by Merck Group (1 mentions) Mip 2, by Thermo Fisher Scientific (1 mentions) Mojosort mouse neutrophil isolation kit, by BioLegend (1 mentions) Annexin 5 apc pi apoptosis detection kit, by Thermo Fisher Scientific (1 mentions) Facs analyzer, by BD (1 mentions) Pvdf membrane, by Thermo Fisher Scientific (1 mentions) Alexa fluor 488, by Cell Signaling Technology (1 mentions) N cadherin, by Cell Signaling Technology (1 mentions) E cadherin, by Cell Signaling Technology (1 mentions) Vimentin, by Cell Signaling Technology (1 mentions) Bx53 microscope, by Olympus (1 mentions)

Close spelling variants of this product

24-well transwell plates NUNC® carrier plate transwell system

17 protocols using transwell plate

1

Chemotaxis Assay of BMDCs

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BMDCs from C57BL/6 or P2X7R-/- mice were added to the upper well of transwell plates (Fisher Scientific, Loughborough, UK) at 1 × 106 per well and 0.1% bovine serum albumin (BSA), CCL5 or CCL20 chemokine (both from R&D Systems) added to the bottom well in a two-fold serial dilution (1000 to 250 pg/ml). After incubating for 3 h at 37 °C, the number of cells in the bottom well were counted in triplicate using a CASY TT cell counter (Roche Innovatis AG, Germany). The percentage of migrated cells was calculated by subtracting background migration from the BSA only data.
Huang S.W., Walker C., Pennock J., Else K., Muller W., Daniels M.J., Pellegrini C., Brough D., Lopez-Castejon G, & Cruickshank S.M. (2016). P2X7 receptor-dependent tuning of gut epithelial responses to infection. Immunology and Cell Biology, 95(2), 178-188.
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2

Chemokine-Induced Transwell Migration

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Colonic lamina propria cells from wild type or Nod2−/− mice were labelled with Vybrant fluorescent dye (Molecular Probes, Leiden, The Netherlands) and added to the upper well of transwell plates (Fisher Scientific, Loughborough, UK) at 1-8×105 per well and chemokines were added to the bottom well (CCL2 (10ng/ml) and CCL2 (1ng/ml), both R and D Systems, Abingdon, UK). As a control, cells were also plated in the absence of chemokine. After incubating for 1h at 37°C cells in the bottom well were stained with CD11c antibodies and the total number of dye-labelled cells counted using an Olympus BX51 upright microscope, captured using a Coolsnap ES camera (Photometrics) through MetaVue Software (Molecular Devices).
Bowcutt R., Bramhall M., Logunova L., Wilson J., Booth C., Carding S.R., Grencis R, & Cruickshank S. (2014). A role for the pattern recognition receptor Nod2 in promoting recruitment of CD103+ Dendritic Cells to the colon in response to Trichuris muris infection. Mucosal immunology, 7(5), 1094-1105.
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3

Chemokine-Induced Transwell Migration

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Colonic lamina propria cells from wild type or Nod2−/− mice were labelled with Vybrant fluorescent dye (Molecular Probes, Leiden, The Netherlands) and added to the upper well of transwell plates (Fisher Scientific, Loughborough, UK) at 1-8×105 per well and chemokines were added to the bottom well (CCL2 (10ng/ml) and CCL2 (1ng/ml), both R and D Systems, Abingdon, UK). As a control, cells were also plated in the absence of chemokine. After incubating for 1h at 37°C cells in the bottom well were stained with CD11c antibodies and the total number of dye-labelled cells counted using an Olympus BX51 upright microscope, captured using a Coolsnap ES camera (Photometrics) through MetaVue Software (Molecular Devices).
Bowcutt R., Bramhall M., Logunova L., Wilson J., Booth C., Carding S.R., Grencis R, & Cruickshank S. (2014). A role for the pattern recognition receptor Nod2 in promoting recruitment of CD103+ Dendritic Cells to the colon in response to Trichuris muris infection. Mucosal immunology, 7(5), 1094-1105.
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4

Chemotaxis Assay for Bone Marrow-Derived Cells

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BMDCs from C57BL/6 or P2X7R−/− mice were added to the upper well of transwell plates (Fisher Scientific, Loughborough, UK) at 1 × 106 per well and 0.1% bovine serum albumin, CCL5 or CCL20 chemokine (both from R&D Systems) added to the bottom well in a twofold serial dilution (1000–250 pg ml−1). After incubating for 3 h at 37 °C, the number of cells in the bottom well were counted in triplicate using a CASY TT Cell Counter (Roche Innovatis AG, Bielefeld, Germany). The percentage of migrated cells was calculated by subtracting background migration from the bovine serum albumin only data.
Huang S.W., Walker C., Pennock J., Else K., Muller W., Daniels M.J., Pellegrini C., Brough D., Lopez-Castejon G, & Cruickshank S.M. (2016). P2X7 receptor-dependent tuning of gut epithelial responses to infection. Immunology and Cell Biology, 95(2), 178-188.
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5

Cell Invasion Assay via Transwell

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To detect cell invasion, transwell chambers were pre-coated with 200 μL of a 0.8 mg/mL Matrigel suspension and dried at 37 ℃. Cells were starved overnight in DMEM medium, 5 × 105 cells in DMEM medium were added to the top chambers of 12-well transwell plates (Nunc; 8 μm pore size), and 15% FBS DMEM medium was added to the bottom chambers. After 48 h incubation, top (non-migrating) cells were removed, and bottom (migrating) cells were fixed with 4% PFA and stained with 5% crystal violet.
Xu J., Galvanetto N., Nie J., Yang Y, & Torre V. (2020). Rac1 Promotes Cell Motility by Controlling Cell Mechanics in Human Glioblastoma. Cancers, 12(6), 1667.
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6

In Vitro Drug Release from Implants

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Implants were embedded in agarose gels in transwell plates (1 implant per insert, membranes: 1.13 cm2, 11 μm, 0.4 μm pore size; Nunc, Roskilde, Denmark), as illustrated in Fig. 1C. The agarose gels were prepared as described above, and the implants included accordingly (placed between 2 “layers” of 0.5 mL gel). The well plates were filled with 4 mL phosphate buffer pH 7.4, covered with lids and Parafilm to minimize evaporation, and placed in a horizontal shaker (80 rpm, 37 °C; GFL 3033). So, again, the total volume of “release medium” (gel + bulk fluid) was 5 mL. At predetermined time points, the entire bulk fluid (4 mL), or 3 mL or 1 mL of the bulk fluid was replaced by fresh (pre-heated) release medium. The withdrawn samples were treated as for the drug release measurements in well agitated bulk fluids.
Bassand C., Benabed L., Freitag J., Verin J., Siepmann F, & Siepmann J. (2022). How bulk fluid renewal can affect in vitro drug release from PLGA implants: Importance of the experimental set-up. International Journal of Pharmaceutics: X, 4, 100131.
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7

Gr1+ Cell-mediated Tumor Priming

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Gr1+ cells were isolated from tumors or spleen at day 23 after injection (see Magnetic cell sorting (MACS) in Supplemental Methods). Afterwards, 1.5 × 105 4T1 or sorted 4T1-SCA1– cells (bottom well) were cocultured in 6-well Transwell plates (0.4 μm, Nunc, ThermoFisher Scientific) with Gr1+CD11b+ sorted cells (top well) at different ratios. After 48 hours of coculture, cells were analyzed for SCA1 expression and the primed cells (4T1 or 4T1-SCA1–) were used for further in vivo or in vitro experiments.
Peyvandi S., Bulliard M., Yilmaz A., Kauzlaric A., Marcone R., Haerri L., Coquoz O., Huang Y.T., Duffey N., Gafner L., Lorusso G., Fournier N., Lan Q, & Rüegg C. (2024). Tumor-educated Gr1+CD11b+ cells drive breast cancer metastasis via OSM/IL-6/JAK–induced cancer cell plasticity. The Journal of Clinical Investigation, 134(6), e166847.
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8

Modulation of Cell Migration by Diabetic Exosomes

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MDA-MB-231 cells were cultured in DMEM media (25 mM glucose + 10% FBS + 1% antibiotic supplementation) for 3 days with adipocyte-derived exosomes from non-diabetic patients (ND group), Type 2 Diabetic patients (T2D group) and fresh media (exosome control) or PBS (control) for three days. Cells were then switched to serum-free media for three hours and subsequently plated in 24-well, 8-micron pore size Transwell plates (Thermo Fisher) for 6 hours. The cells were plated in the upper well of the transwell inserts with serum free media and the bottom well was filled with DMEM complete media to serve as a chemoattractant. Cells that stayed in the upper side of the membrane and did not migrate by the end of the assay were removed with a cotton swab, and the cells that migrated were fixed with ice-cold methanol for 5 minutes at −20°C. After fixation, cells were then stained with 1% crystal violet (v:v) in 2% ethanol for 10 minutes at room temperature. Images were captured by an EVOS XL Core digital inverted microscope. The percentage of migration and invasion was determined by first calculating the sum of the area of total migrated/invaded cells on the entire membrane with ImageJ software (National Institutes of Health, Bethesda, MD), and then converted to relative percent migration/invasion by comparing each condition to the control condition.
Jafari N., Kolla M., Meshulam T., Shafran J.S., Qiu Y., Casey A.N., Pompa I.R., Ennis C.S., Mazzeo C.S., Rabhi N., Farmer S.R, & Denis G.V. (2021). Adipocyte-derived exosomes may promote breast cancer progression in type 2 diabetes. Science signaling, 14(710), eabj2807.
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9

In Vitro Endothelial Permeability Assay

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Brain endothelioma bEnd.3 cells were cultured in DMEM, supplemented with 10% FBS, and harvested by detachment with TrypLE (Thermo Fisher Scientific). For in vitro permeability assay, bEnd.3 cells or isolated primary endothelial cells were seeded on fibronectin-coated (7 µg/mL, Sigma-Aldrich) or collagen-precoated Transwell plates (0.4 µm pore size; Thermo Fisher Scientific) and cultured for 3 d. Cells were then incubated with 5 µM corticosterone (Sigma-Aldrich) and 1 mg/mL 40-kDa FITC-dextran (Sigma-Aldrich) for 20 h. Medium from the receiver tray was collected, diluted to 1:20 in PBS, and fluorescence was measured with a spectrophotometer (Perkin-Elmer) at an excitation wavelength of 490 nm and emission of 529 nm.
Na S.Y., Janakiraman M., Leliavski A, & Krishnamoorthy G. (2021). High-salt diet suppresses autoimmune demyelination by regulating the blood–brain barrier permeability. Proceedings of the National Academy of Sciences of the United States of America, 118(12), e2025944118.
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10

Neutrophil Migration Assay in Mice

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Bone marrow neutrophils were isolated from Wt and α2BAR-Tg mice using a MojoSortTM Mouse Neutrophil Isolation Kit (Biolegend, US). The isolated α2BAR-Tg and Wt neutrophils were stained with CellTrace™ Violet (V440) or CellTrace™ CFSE, respectively. Labeled cells were mixed at the at the ratio of 1:1 and then adoptively transferred into MSU-pretreated (3 mg/per mouse) Wt or α2BAR-Tg mice. Splenocytes and PECs were harvested after 6 h. The ratio and numbers of Wt (CFSE) and α2BAR-Tg (V440) cells in the spleen and peritoneal fluid were determined by flow cytometry. For the in vitro neutrophil migration, the mixed neutrophils in Dulbecco modified Eagle medium with 10% Fetal Bovine Serum were placed in the insert of a 6-μm Transwell® plate (Thermo Scientific) at the concentration of 1 × 106 cells/mL (100 μl/well). The lower chamber contained 600 ul Dulbecco modified Eagle medium with 10% Fetal Bovine Serum and MIP-2(200 ng/ml, PeproTech). After incubation for 4 h in 37°C, the cells in the insert and bottom wells were collected and the ratio of Wt (CFSE) vs. α2BAR-Tg (V440) cells were detected by fluorescence microscope and flow cytometry.
Duan L., Chen J., Razavi M., Wei Y., Tao Y., Rao X, & Zhong J. (2019). Alpha2B-Adrenergic Receptor Regulates Neutrophil Recruitment in MSU-Induced Peritoneal Inflammation. Frontiers in Immunology, 10, 501.
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