Matrigel coated film insert

Manufactured by BD

Sourced in United States

Matrigel-coated film insert is a laboratory product designed for cell culture applications. It consists of a thin film coated with a mixture of extracellular matrix proteins derived from Engelbreth-Holm-Swarm (EHS) mouse sarcoma cells, commonly referred to as Matrigel. This product provides a biologically active substrate for the attachment, growth, and differentiation of various cell types.

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Lab products found in correlation

24 well invasion chambers, by BD (4 mentions) Fluorescence microscope, by Olympus (2 mentions) Calcein am, by Thermo Fisher Scientific (1 mentions)

Spelling variants of this product

Matrigel (17210 citations) Matrigel-coated (225 citations) Matrigel-coated inserts (53 citations) Matrigel film (4 citations)

4 protocols using matrigel coated film insert

Matrigel-Based Cell Invasion Assay

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Cell invasion was performed using the Matrigel-coated film insert (8 mm pore size) fitted into 24-well invasion chambers (BD Biosciences, San Jose, CA, USA) [44 (link)]. After 24 h incubation, the filter inserts were removed from the wells and the cells on the top surface of the filter were removed using cotton swabs. The cells on the bottom surface of the filter were stained with 4′, 6-diamidino-2-phenylindole (DAPI) for 1 min, washed 3 times with PBS, and the cell number was counted under an Olympus fluorescence microscope.

Yang C.J., Liu Y.P., Dai H.Y., Shiue Y.L., Tsai C.J., Huang M.S, & Yeh Y.T. (2015). Nuclear HDAC6 inhibits invasion by suppressing NF-κB/MMP2 and is inversely correlated with metastasis of non-small cell lung cancer. Oncotarget, 6(30), 30263-30276.

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Measuring Cell Invasion Using Matrigel

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Cell invasion was measured using 24-well invasion chambers (BD Biosciences; San Jose, CA) with the Matrigel-coated film insert (8 μm pore size) as described previously [7 (link)]. In brief, A2058 cells (5 × 10⁴), which were resuspended in serum-free DMEM supplemented with 0.1% BSA, were added to the top compartment of the invasion chamber. The various compounds were preincubated in serum-free DMEM/0.1% BSA with recombinant ATX for 30 min at 37°C, followed by the addition of 1 μM LPC 18:1 to the bottom chamber. The invasion chambers were incubated at 37°C in 5% CO₂ for 16 h. Next, the filter inserts were removed from the wells and transferred to a new 24-well plate containing 4 μg/ml of calcein AM (Molecular Probes, Invitrogen) in Hank’s balanced salt solution. After a 1-h incubation, the fluorescence of invaded cells was measured with a FLEXStation 3 plate reader at excitation and emission wavelengths of 485 and 530 nm, respectively.

Fells J.I., Lee S.C., Norman D.D., Tsukahara R., Kirby R.J., Nelson S., Seibel W., Papoian R., Patil R., Miller D.D., Parrill A.L., Pham T.C., Baker D.L., Bittman R, & Tigyi G. (2014). Targeting the Hydrophobic Pocket of Autotaxin with Virtual Screening of Inhibitors Identifies a Common Aromatic Sulfonamide Structural Motif. The FEBS journal, 281(4), 1017-1028.

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Cell Migration and Invasion Assay

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Cells were seeded in six-well plates, scratched with 10 µl pipet tips and cultured in 1× DMEM solution without serum. Then, the cells were washed three times with phosphate-buffered saline (PBS) and photographed by microscopy (Leica, Wetzlar, Germany) at 0 hours, 6 hours and 12 hours. The migration distance was measured by Image-Pro Plus software.
Cells were seeded in 24-well invasion chambers (BD Biosciences, New Jersey, USA) with a Matrigel-coated film insert (8 mm pore). The mixed solution was diluted to generate a 1× DMEM solution containing 10% serum. Two days later, cells on the bottom surface of the filter were subjected to staining with crystal violet for 15 min and then washed three times with PBS, and the cell number was counted under a microscope (Leica, Wetzlar, Germany).

Li J., Chen X., Zhu L., Lao Z., Zhou T., Zang L., Ge W., Jiang M., Xu J., Cao Y., Du S., Yu Y., Fan G, & Wang H. (2021). SOX9 is a critical regulator of TSPAN8-mediated metastasis in pancreatic cancer. Oncogene, 40(30), 4884-4893.

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Quantifying Cell Invasion in Response to EGF

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Cells were seeded in 24-well invasion chambers (BD Biosciences, San Jose, CA, USA) with the Matrigel-coated film insert (8 mm pore). The mixed solution was diluted to give a 1× DMEM solution containing 10% serum. The cells were seeded in absence or presence of EGF (100 ng/ml) (chemokinesis). Two days later, cells on the bottom surface of the filter subjected to staining with 4′,6-diamidino-2-phenylindole for 1 min, then washed three times with PBS, and the cell number was counted under a fluorescence microscope (Olympus).

Chen T., Li J., Xu M., Zhao Q., Hou Y., Yao L., Zhong Y., Chou P.C., Zhang W., Zhou P, & Jiang Y. (2017). PKCε phosphorylates MIIP and promotes colorectal cancer metastasis through inhibition of RelA deacetylation. Nature Communications, 8, 939.

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