Transwellfilters with 8 μm pores

Manufactured by Corning

Sourced in United States

Transwell filters with 8 μm pores are a type of laboratory equipment used for cell culture applications. They consist of a permeable membrane with pores that allow the passage of cells, molecules, or other materials between the upper and lower chambers of the device. The 8 μm pore size is suitable for various cell types and applications that require controlled migration or co-culture experiments.

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Lab products found in correlation

Calcein, by Thermo Fisher Scientific (1 mentions) Gelatin, by Merck Group (1 mentions) Ultrapure flagellin, by InvivoGen (1 mentions) Eclipse 80i microscope, by Nikon (1 mentions)

Close spelling variants of this product

8-μm pores

4 protocols using transwellfilters with 8 μm pores

Assessing Migration of Glioblastoma Cells

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To assess the migration ability of
U87 MG and primary GBM cells after treatment with TMZ and amlexanox,
alone or in combination, for 48 h, a scratch wound healing assay was
conducted. In brief, the treated cells (3 × 10⁵ cells/well)
were seeded into six-well plates; when the cells reached 80–90%
confluence in a monolayer, scratch wounds were made using a 200 μL
pipette tip. Then, cell debris was removed, and a microscope was used
to confirm the uniform scratch width of every group. After incubation
at 0, 12, and 24 h, five different fields of each well were measured
and photographed using a phase-contrast microscope. In addition, transwell
filters with 8 μm pores (Corning Costar, NY) (without Matrigel)
were also used to evaluate the migration ability of treated cells.
The assay was conducted as described in a previous report.^{21 (link)}

Xiong J., Guo G., Guo L., Wang Z., Chen Z., Nan Y., Cao Y., Li R., Yang X., Dong J., Jin X., Yang W, & Huang Q. (2021). Amlexanox Enhances Temozolomide-Induced Antitumor Effects in Human Glioblastoma Cells by Inhibiting IKBKE and the Akt-mTOR Signaling Pathway. ACS Omega, 6(6), 4289-4299.

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Leukocyte-Endothelial Adhesion and Transmigration Assay

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For the leukocyte–endothelial adhesion assay, confluent HUVEC monolayers were pretreated with TNF or IL-1β after the overexpression of adenoviral SENP1-Mut, various GATA2 constructs or vector control. Calcein (Molecular Probes)-labelled THP-1 monocytes were co-cultured with stimulated HUVECs for 60 min, gently washed twice to remove non-adherent cells and microphotographs were acquired. At least five random fields were quantified per condition.
The transmigration assay was performed using 6.5 mm Transwell filters with 8-μm pores (Costar). HUVECs were seeded onto the insert, which was coated with 0.1% gelatin (Sigma-Aldrich), and cultured until confluent. HUVEC monolayers were stimulated with TNF or IL-1β for 24 h before Calcein-labelled THP-1 was added into the chamber. After incubating for 24 h, the number of cells that transmigrated to the bottom compartment was imaged and quantified.

Qiu C., Wang Y., Zhao H., Qin L., Shi Y., Zhu X., Song L., Zhou X., Chen J., Zhou H., Zhang H., Tellides G., Min W, & Yu L. (2017). The critical role of SENP1-mediated GATA2 deSUMOylation in promoting endothelial activation in graft arteriosclerosis. Nature Communications, 8, 15426.

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Chemotaxis Assay for Smooth Muscle Cells

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Harvested wild-type and Nox4 knockout SMCs (8 × 10³ cells) were replated onto the upper chambers of Transwell filters with 8 μm pores (Corning, NY, USA) coated with 10 μg/mL fibronectin, and the chambers were placed in serum-free DMEM with or without 100 ng/mL ultrapure flagellin (InvivoGen, #tlrl-epstfla, San Diego, CA, USA). After 16 h, the cells were fixed with 70% methanol for 5 min. The cells on the underside of each filter were stained with hematoxylin for 5 min and rinsed with distilled water. The filter was stained with eosin for 3 min and washed again. Nonmigrating cells on the upper side of the filter were removed with cotton swabs. Images were captured using a Nikon Eclipse 80i microscope. Three independent filters were analyzed for each experiment by counting the number of cells on each filter in five random fields.

Kim J., Yoo J.Y., Suh J.M., Park S., Kang D., Jo H, & Bae Y.S. (2019). The flagellin-TLR5-Nox4 axis promotes the migration of smooth muscle cells in atherosclerosis. Experimental & Molecular Medicine, 51(7), 78.

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Quantifying Cell Migration Using Transwell Assay

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Cell migration was measured in Transwell filters with 8-μm pores (Corning, Tewksbury, MA, USA). Inserts were coated with human fibronectin protein (Cat. #33016015, Thermo Fisher Scientific, Waltham, MA, USA). Sorted cells (2 × 10⁴ cells), in 200 μL of medium, were loaded into the upper chambers, and 500 μL of the hOPC medium was added to the chamber. After incubation for 18 h at 37 °C, a cotton swab was used to wipe the cells that had not migrated onto the upper surface of the chamber. The migrated cells were fixed with 4% paraformaldehyde and stained with DAPI (1:20). Images were acquired using a fluorescence microscope. The migrated cells were quantified using ImageJ software [24 (link)] by analysing cell nuclei from at least five randomly selected fields per chamber. The cell migration rate was calculated as follows: cell migration rate (V1%) = N1/N2 × 100%, where N1 refers to the initial cell seeding number, and N2 refers to the number of migrated cells. The experiment was repeated three times independently in the laboratory.

Zhou H., He Y., Wang Z., Wang Q., Hu C., Wang X., Lu S., Li K., Yang Y, & Luan Z. (2021). Identifying the functions of two biomarkers in human oligodendrocyte progenitor cell development. Journal of Translational Medicine, 19, 188.

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