Cell invasive activity was estimated using transwell plates coated with extracellular matrix gel from Corning (Corning, NY, United States). Cells (1 × 105) were plated in the upper chambers containing 100 μL serum-free medium, whereas 500 μL medium containing 10% FBS was loaded into the lower chamber. After 24 h of incubation, the cells that migrated to the lower membrane surface were fixed with 4% paraformaldehyde and then stained with 0.1% crystal violet. The number of migratory cells was observed and counted under a light microscope. For the wound-healing migration assay, after the cells cultured in a 6-well plate reached 90% confluency, wounds were created by scraping the bottom of the wells with a 200 μL yellow plastic pipette tip. The wounds were observed and photographed at 0 h and 48 h.
Extracellular matrix gel
Manufactured by Corning
Sourced in United States
Extracellular matrix gel is a laboratory product designed to mimic the natural environment of cells. It provides a three-dimensional substrate that supports the growth and behavior of cells in in vitro cell culture experiments.
Automatically generated - may contain errors
Lab products found in correlation
9 protocols using extracellular matrix gel
1
Cell Invasion and Migration Assay
2
Xenograft model of neuroblastoma
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Female BALB/c nude mice (4 weeks old) received a subcutaneous injection of NB cells (2 × 106 cells per mouse) suspended in 100 μl of extracellular matrix gel (Corning, NY, United States) into their front flanks. The mice were randomly divided into two groups (n = 5) and treated with CYN once a day for 3 weeks (0 or 5 mg/kg, i.g.). This concentration was referred to the article by Zhang et al. (Zheng et al., 2020 (link)). Tumour volumes and mouse weights were measured every 3 days. Tumour volumes were calculated with the formula (length × width × height)/2. After 3 weeks, all the mice were sacrificed by euthanasia, and afterwards, the tumours were quickly collected for weight measurements and then embedded in paraffin for immunohistochemistry.
3
Transwell Invasion Assay for NSCLC
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The invasive activity of NSCLC cells was estimated using transwells (8 μm pore size, 6.5 mm in diameter, polycarbonate membrane) coated with extracellular matrix gel (Corning, Tewksbury, MA, USA). An aliquot of 1 × 105 cells was placed in the upper chamber with 0.1 mL serum‐free medium, while the lower chamber was loaded with 0.5 mL of medium containing 10% fetal bovine serum. After incubation for 24 hours, the cells were fixed with 4% paraformaldehyde and then counterstained with 0.1% crystal violet. The cells that had migrated into the lower chamber were observed and counted under a light microscope.
4
Transwell Migration and Invasion Assay
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Cells were trypsinized and collected in respective 2% serum media and pelleted at 1000 rpm for 5 minutes. Cells were resuspended in respective 2% serum media and counted using the countess automated cell counter and tryphan blue (Invitrogen, Carlsbad, USA). For the transwell migration assay, 2 × 104/200μl H446 or H196 cells were plated in the top chamber with the non-coated membrane (8 μm for 24-well plate; Corning Costar, NY, USA). For the invasion assay, 5×104/200 μl H446 or H196 cells were plated in the top chamber with extracellular matrix gel (Corning, USA)-coated membranes (8 μm for 24-well plate; Corning Costar, NY, USA). In both assays, cells that did not migrate through or invade the pores were removed with a cotton swab. Cells on the lower surface of the membrane were fixed with methanol and stained with 0.1% crystal violet (Sigma, USA). Experiments were performed in three times. The invaded cells were photographed with an inverted phase-contrast microscope (Olympus, CKX41), and counted at least 3 randomly-selected images to represent the relative migration and invasion. H196 were counted by the average number of cells and H446 were counted by relative area per field of view.
5
HeLa 3D Cell Migration and Invasion Assay
6
Quantifying Cell Migration Potential
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The invasive activity of HPCs was estimated using transwells (6.5 mm in diameter, polycarbonate membrane, 8 μm pore size) coated with extracellular matrix gel obtained from Corning Inc. (Corning, NY, USA). After 24 h of transfection, an aliquot of 1 × 105 cells was placed in the upper chamber with 0.1 ml serum-free medium, whereas the lower chamber (24-well plate) was loaded with 0.5 ml of medium containing 10% fetal bovine serum. After 24 h of incubation, the cells were fixed with 4% paraformaldehyde and then counterstained with 0.1% crystal violet. The cells that had migrated into the lower chamber were observed and counted under a light microscope. Then the number of migratory cells was calculated.
7
Extracellular Matrix Invasion Assay
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The upper chambers were coated with 50 μg extracellular matrix gel (Corning, Corning, NY, USA). After incubation for 24–48 h, the invaded cells on the lower membrane were stained with 4% paraformaldehyde, then stained with 0.1% crystal violet. The upper cells were moved gently by the soft medical cotton ball. For each group, 10 pictures were randomly taken under a microscope with 400× magnification (Olympus, Tokyo, Japan). Cell numbers per field were calculated by ImageJ.
8
Cell Migration Assay
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An aliquot of 1×105 cells in 0.1 mL serum-free medium was placed in the upper chamber of 6.5 mm, 8 µm pore size polycarbonate membrane that was precoated with extracellular matrix gel (Corning, NY, USA). The lower chamber was loaded with 0.5 mL of medium containing 20% fetal bovine serum. The cells were fixed with 10% paraformaldehyde after 48 h incubation and then counterstained with 0.1% crystal violet. Cells migrating to the lower chamber were observed by light microscopy, and the number of migrating cells was calculated.
9
Cell Migration and Invasion Assay
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For the migration assay, 4 × 10 4 Bel7402 cells in serum-free medium were seed in the upper compartment of an 8 µm transwell chamber (Corning, NY, USA). For the invasion assay, the upper chambers were previously coated with 50 µg extracellular matrix gel (Corning, NY, USA). After incubation for 24-48 hours, the migrated or invaded cells on the lower membrane were stained with 4% paraformaldehyde, then stained with 0.1% crystal violet. The upper cells were moved gently by the soft medical cotton ball. Each group random took 10 pictures under a microscope with 400× magni cation (Olympus, Japan). Cell numbers per eld were calculated by ImageJ.
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