24 well transwell

Manufactured by Corning

Sourced in United States, China

The 24-well transwell is a laboratory equipment used for cell culture studies. It consists of a upper and lower chamber separated by a porous membrane, allowing for the study of cell migration, permeability, and co-culture experiments.

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Lab products found in correlation

Matrigel, by BD (28 mentions) Matrigel, by Corning (8 mentions) Inverted microscope, by Olympus (3 mentions) Crystal violet, by Merck Group (3 mentions) Image pro plus 6, by Media Cybernetics (2 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions) Cell counting kit 8 (cck8), by Dojindo Laboratories (2 mentions) Lsm 880, by Zeiss (2 mentions) Cxcl12, by Thermo Fisher Scientific (1 mentions) Carboxyfluorescein succinimidyl ester (cfse), by Merck Group (1 mentions) Application suite, by Leica (1 mentions) Hct 8 cells, by American Type Culture Collection (1 mentions) Dm2500, by Leica (1 mentions) Pf573228, by Merck Group (1 mentions) Ly294002, by Merck Group (1 mentions) Amd3100, by Merck Group (1 mentions) Cytoselect 24 well cell invasion assay kit, by Cell Biolabs (1 mentions) Versamax microplate reader, by Molecular Devices (1 mentions) Thiazolyl blue tetrazolium bromide mtt, by Beyotime (1 mentions) Paraformaldehyde fix solution, by Beyotime (1 mentions)

Close spelling variants of this product

24-well transwell plates (425 citations) Transwell chambers (24-well insert; (9 citations) 24-well transwell cell culture chambers (8 citations) 24-well plate transwell chamber (6 citations) HTS Transwell 24 wells (5 citations) Costar 24-well transwell plates (5 citations) 24-well plate transwells (4 citations) 24-well Transwell assay (4 citations) 24-transwell Boyden chamber wells (3 citations) 24 wells (2 citations)

84 protocols using 24 well transwell

Transwell Chemotaxis and Transmigration Assays

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Chemotaxis assays were carried out in 6.5-mm diameter/8.0-μm pore size 24-well transwells (Costar, Corning, NY, USA). Control Mock, L-Eng and S-Eng U937 transfectants were CFSE-labelled and serum-starved 24 hours before the assay. Then, 5x10 4 cells in 100 μL were loaded into the upper chamber and chemotaxis was carried out for 3 hours at 37ºC to the lower chamber with or without 100 ng/mL recombinant human MCP-1 (CCL2). As a positive control, recombinant SDF-1α (CXCL12) was assayed at 100 ng/mL.
For transmigration assays through an endothelial monolayer, 5x10 4 HUVECs were seeded onto a gelatin pre-coated upper chamber and allowed to form a complete monolayer on the porous membrane of 6.5-mm diameter/5.0-μm pore size 24-well transwells (Costar). U937 transfectants were CFSE-labelled and assayed by adding or not recombinant human MCP-1 to the lower chamber as a chemoattractant at 100 ng/mL for 4 hours. Viable migrated cells in the lower chambers were collected, washed with PBS and counted in the flow cytometer by analysing each sample in the same predetermined time and flow conditions.

Blanco F.J., Ojeda-Fernandez L., Aristorena M., Gallardo-Vara E., Benguria A., Dopazo A., Langa C., Botella L.M, & Bernabeu C. (2015). Genome-wide transcriptional and functional analysis of endoglin isoforms in the human promonocytic cell line U937. Journal of cellular physiology, 230(4).

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Assessing Glioma Cell Migration and Invasion

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The in vitro migratory and invasive ability of glioma cells was assessed using the transwell chamber method [25 (link)]. In brief, U251 glioma cells were seeded into 24-well transwells (Corning Corp. USA) at a density of 5 × 10⁵ cells in 200 μL of medium in the upper chamber and were incubated in serum-free medium with or without melatonin, and the bottom chamber was filled with 600 μL of medium containing 10% FBS. After 24 hours of incubation at 37°C, non-migrating cells on the upper surface of the membrane were scrubbed gently with a cotton-tipped swab. The migratory cells on the lower surface of the membrane were fixed with 95% methanol and stained with 0.1% crystal violet (Sigma-Aldrich, MO, USA). Stained migratory cells were photographed under an inverted light microscope and quantified by manual counting in five randomly selected areas of view. Five independent experiments were performed. For invasion assays, U251 glioma cells were seeded into diluted matrigel-precoated 24-well transwells (Corning Corp. USA) at a density of 5 × 10⁵ cells in 200 μL of medium in the upper chamber. The procedure of cell treatment and staining was similar with transwell migration assay.

Xu C.S., Wang Z.F., Huang X.D., Dai L.M., Cao C.J, & Li Z.Q. (2015). Involvement of ROS-alpha v beta 3 integrin-FAK/Pyk2 in the inhibitory effect of melatonin on U251 glioma cell migration and invasion under hypoxia. Journal of Translational Medicine, 13, 95.

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Mesenchymal Stem Cell Exosome Effects on Leukocyte Chemotaxis

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We examined the effect of MSC-Exo on the chemotaxis of leukocytesin vitro. In this experiment, the indicator cells were splenocytes prepared from immunized rat on day 12 post immunization. The cells were added to the upper well of a microchemotaxis device (5 μm pore size, 24wellTranswell; Corning-Costar), and RPMI 1640 medium containing chemokine (C-C motif) ligand 2 and 21 (10 nMmacrophage chemoattractant CCL2; 200 ng/ml lymphocyte chemoattractant CCL21; BioLegend) were added to the lower well, with or without MSC-Exo (10 ug/ml). The cells that migrated to the lower well after 4 h were collected and the number of CD4⁺, CD161⁺, CD68⁺ and Gr⁺ cells were determined by flow cytometry. All assays were performed in triplicate.

Bai L., Shao H., Wang H., Zhang Z., Su C., Dong L., Yu B., Chen X., Li X, & Zhang X. (2017). Effects of Mesenchymal Stem Cell-Derived Exosomes on Experimental Autoimmune Uveitis. Scientific Reports, 7, 4323.

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Matrigel Invasion Assay for PC Cells

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After transfection, PC cells were resuspended in serum-free DMEM and plated into the upper chamber of 24-well transwell (pore size 8 µm; Corning Costar) coated with Matrigel (BD Biosciences). The lower chamber was filled with DMEM containing 10% FBS. After incubation for 24 h, noninvaded cells were removed with cotton swabs, while invaded cells were fixed with 4% paraformaldehyde, and then stained with 0.1% crystal violet for 20 min. The invaded cells were imaged with an inverted microscope (Olympus).

Zhang J., Shen Y., Ma D., Li Z., Zhang Z, & Jin W. (2022). SLCO4A1-AS1 mediates pancreatic cancer development via miR-4673/KIF21B axis. Open Medicine, 17(1), 253-265.

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Cytotoxicity of CCN@E. coli on 4T1 Cells

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The MTT assay was used to measure the cytotoxicity of CCN@E. coli. 4T1 cells were seeded under the down chamber of 24-well transwell® (Corning Costar, USA) filters with a density of 5 × 10⁴ cells per well. CCN@E. coli was placed on the up chamber of the assay at a density of 1 × 10⁶, 1 × 10⁷, and 1 × 10⁸ CFU mL⁻¹ (100 μL each well). Four hours after the irradiation ( > 630 nm, 30 mW cm⁻², 15 min), the up chamber was removed. Twenty-four hours later, MTT (50 μL, 5 mg mL⁻¹) was added into each well and co-incubated for another 4 h. The cultured medium in each well was replaced with dimethylsulfoxide (DMSO) (750 μL). The absorbance at 570 nm was measured with SpectraMax i3x multi-mode detection platform. The relative cell viability was counted according to the following formula: cell viability (%) = (OD_{570 sample} − OD_{570 blank}) / (OD_{570 control} – OD_{570 blank}) × 100%.

Zheng D.W., Chen Y., Li Z.H., Xu L., Li C.X., Li B., Fan J.X., Cheng S.X, & Zhang X.Z. (2018). Optically-controlled bacterial metabolite for cancer therapy. Nature Communications, 9, 1680.

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Chemotaxis Assay for CXCL12

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Transfected cells (1×10⁵) labeled with CFSE (Sigma) were seeded in the upper chamber of 24-well transwell (5 µm pore size; Costar). Complete medium with or without CXCL12 (100 ng/ml; peprotech) was added in the lower chamber. Three hours later, cells in the lower chamber were evaluated by fluorescent microscopy (OLYMPUS DP72) and counted. Chemotaxis index was calculated as the ratio of cells migrating toward CXCL12 to cells migrating toward the control medium.

Liu J., Li W., Wang S., Wu Y., Li Z., Wang W., Liu R., Ou J., Zhang C, & Wang S. (2014). MiR-142-3p Attenuates the Migration of CD4+ T Cells through Regulating Actin Cytoskeleton via RAC1 and ROCK2 in Arteriosclerosis Obliterans. PLoS ONE, 9(4), e95514.

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Transwell Permeability Assay for OGD/R

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Cells (1 × 10⁵) were seeded into the upper chamber of polyester membranes in a 24-well Transwell (pore size 0.4 μm; diameter 6.5 mm; Costar, NY) and cultured in 0.5 ml of DMEM. Medium (1.5 ml) was added to the lower chamber, and Transwells were incubated for 2 days until the cells reached 100% confluence. The cells were subjected to the OGD/R procedure described above, and at 1 h before the end of the reoxygenation period, FITC-dextran solution was added to the upper chamber of each Transwell. Sample readings were converted to FITC-dextran concentrations based on a standard curve. The permeability coefficient (Pc) of FITC-dextran was described as Pc = [A]/t × 1/A × V/[L], where [A] denotes the concentration of FITC-dextran in the lower chamber; [L], the concentration in the upper chamber; A, the area of the membrane (cm²); t, time (s); and V, the volume of the lower chamber (Zhang et al., 2015 (link)).

Wang T., Liu C., Pan L.H., Liu Z., Li C.L., Lin J.Y., He Y., Xiao J.Y., Wu S., Qin Y., Li Z, & Lin F. (2020). Inhibition of p38 MAPK Mitigates Lung Ischemia Reperfusion Injury by Reducing Blood–Air Barrier Hyperpermeability. Frontiers in Pharmacology, 11, 569251.

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Coculture of NK Cells and SH-SY5Y Cells

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For coculture experiments in transwell condition, 2 × 10⁵ resting NK cells were cultured for 48 h with 3 × 10⁵ SH-SY5Y cells. NK cells and SH-SY5Y cells were placed in 24-well transwell (0.3-µm pore size; Corning Costar), upper and bottom chamber, respectively (12 (link)).

Regis S., Caliendo F., Dondero A., Casu B., Romano F., Loiacono F., Moretta A., Bottino C, & Castriconi R. (2017). TGF-β1 Downregulates the Expression of CX3CR1 by Inducing miR-27a-5p in Primary Human NK Cells. Frontiers in Immunology, 8, 868.

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Angiogenic Factors Modulate HUVEC Migration

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2.5×10⁵ T98, U118, U87MG cells that were genetically modified to vary in EMP2 expression were cultured in 6-well plates for 48 hours. Conditioned media were collected at the end of incubation. To determine the biologic effects of cell-secreted angiogenic factors into the conditioned media, 1×10⁴ HUVEC cells were plated in the top chamber inserts of 24-well transwell (Corning Costar, Tewksbury, MA) and conditioned media from the cells were added into the bottom wells and the co-culture was incubated at 37°C for 5 hours. The filters of inserts were fixed with 3.7% formaldehyde at room temperature for 10 minutes and followed by staining with 0.1% crystal violet in 20% methanol at room temperature for 10 min. The migrated HUVEC cells were visualized using bright field microscopy. Migratory cell numbers were averaged by counting four random fields per transwell under 40× magnification. The experiment was repeated at least three times.
To determine the effect of the anti-EMP2 IgG1 antibody on cell-secreted angiogenic factors, 2.5×10⁵ U87MG wild type cells were cultured in 6-well plates in the presence of 50 µg/ml anti-EMP2 full-length IgG1 antibody (n=4) or a vehicle control (n=3) for 48 hours. Conditioned media were collected at the end of incubation. HUVEC migration assay was performed as described above.

Qin Y., Takahashi M., Sheets K., Soto H., Tsui J., Pelargos P., Antonios J.P., Kasahara N., Yang I., Prins R.M., Braun J., Gordon L.K, & Wadehra M. (2017). Epithelial Membrane Protein-2 (EMP2) Promotes Angiogenesis in Glioblastoma Multiforme. Journal of neuro-oncology, 134(1), 29-40.

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Quantitative Cell Invasion Assay

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Cell invasion assay was performed in a 24-well Transwell™ (Costar). The upper chamber surface of the filter was coated with Matrigel (Corning) before the experiment. The cells were prepared (1 × 10⁵/200 μl) with serum-free DMEM and loaded into the upper chamber. DMEM medium containing 20% FBS was added to the bottom chamber as the chemoattractant. After 24 h incubation, wet cotton was used to remove the non-invaded cells from the wells. The cells were fixed with carbinol for 15 min at room temperature, stained with 0.1% crystal violet for 20 min, and quantified by counting the total number of cells in four independent areas under the ZEISS Axio Imager.Z2 Microscope.

Yang F., Jin H., Que B., Chao Y., Zhang H., Ying X., Zhou Z., Yuan Z., Su J., Wu B., Zhang W., Qi D., Chen D., Min W., Lin S, & Ji W. (2019). Dynamic m6A mRNA methylation reveals the role of METTL3-m6A-CDCP1 signaling axis in chemical carcinogenesis. Oncogene, 38(24), 4755-4772.

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