To assay the invasive capacity of tumor cells, Transwell migration assays were carried out by using 24-well MILLI cell Hanging Cell Culture inserts 8 mm PET (MILLIPORE) coated with Matrigel matrix gel (BD Biosciences) according to the manufacturer’s protocol. After transfection for 48 h, cells suspended in serum-free medium were added into the upper chamber. After 24 h incubation, cells on the underside of the membrane were fixed with 90% alcohol and stained with 0.1% crystal violet solution. The values for invasion were obtained by counting 6 fields per membrane.
Millicell hanging cell culture inserts 8 mm pet
Manufactured by Merck Group
Sourced in United States
The MILLIcell Hanging Cell Culture inserts 8 mm PET are laboratory equipment used for in vitro cell culture applications. These inserts are made of polyethylene terephthalate (PET) and have a diameter of 8 millimeters. They are designed to be suspended in a cell culture medium to facilitate the growth and study of cells.
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4 protocols using millicell hanging cell culture inserts 8 mm pet
1
Transwell Invasion Assay for Tumor Cells
2
Transwell Invasion Assay for Glioma Cell Migration
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The transwell migration assay was performed using 24-well MILLI Cell Hanging Cell Culture inserts 8 mm PET (Millipore, Bedford, MA, USA) as previously described27 (link). The cells were cultured in DYT-40 (2.5, 5 and 10 μM) for 24 h, and in parallel, pcDNA3.1-AEG-1 was transfected into U251 and U87 cells for 24 h and treated with DYT-40 (10 μM) for an additional 24 h. Subsequently, the cells were seeded in serum-free medium onto triplicate wells of BD BioCoatTM MatrigelTM Invasion Chambers (BD Bioscience), and complete medium containing 10% fetal bovine serum was added to the lower chamber. The cells invaded through the polycarbonate membrane were stained using crystal violet after incubation for 24 h. Image magnification: 200×. Five random fields from each of the triplicate invasion assays were counted using phase-contrast microscopy.
3
Transwell Migration and Invasion Assay
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Transwell migration and invasion assays were performed by 24-well MILLIcell Hanging Cell Culture inserts 8 mm PET (MILLIPORE) precoated with or without Matrigel matrix gel (BD Biosciences). The detailed procedure was referred to the previous study.7 A total of 8×104 cells/200 µL in serum-free medium were added to the upper chamber. Twenty percent FBS in 800 µL medium was used as a chemoattractant in the bottom chamber. Cells were allowed to migrate or invade for 24 h. Cells on the upper side of the membrane were wiped off, and cells on the lower side of the membrane were fixed and stained with crystal violet solution. Five random fields from each of the triplicate migration assays were counted by using phase contrast microscopy. Quantification was done by measuring with Microplate Reader (optical density 570 nm) after being destained with glacial acetic acid. All migration and invasion assays were done in triplicate for at least three independent experiments.
4
Transwell Invasion Assay for Cancer Cells
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The would healing assay was performed to examine the migration ability of cancer cells. The detailed procedure was referred to the previous study [19] . Transwell invasion assay was carried out by using 24-well MILLIcell Hanging Cell Culture inserts 8 mm PET (MILLIPORE) pre-coated with Matrigel matrix gel (BD Biosciences). Briefly, a total of 8 -10 × 10 4 cells/200 µl in serum-free medium were added to the upper chamber. Twenty percentage FBS in 800 µl medium was used as a chemo-attractant in the bottom chamber. Cells were allowed to invade for 24 h. Cells on the underside were fixed in methanol and stained with 0.1 % crystal violet solution. Five random fields from each of the triplicate migration assays were counted by using phase contrast microscopy. Quantification was done by measuring with Microplate Reader (OD 570 nm) after being destained with glacial acetic acid.
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