GH3 cells (5 × 105 per well) were seeded in the upper chambers in 24-well culture plates with 8-μm pores (Corning, USA). GH3 cells were allowed to migrate through the pores in the transwell membrane during incubation at 37 °C with 5% CO2 for 24 h. Then, cells on the lower surface of the membrane were fixed in 4% paraformaldehyde for 15 min, stained with crystal violet for 10 min, washed with PBS. The migrated GH3 cells were imaged under a microscope and counted using the ImageJ software.
8 μm pores
Manufactured by Corning
Sourced in United States
The 8-μm pores are a laboratory equipment product designed for various applications. These pores have a diameter of 8 micrometers and are suitable for various filtration and separation processes in research and scientific settings.
Automatically generated - may contain errors
5 protocols using 8 μm pores
1
Transwell Migration Assay for GH3 Cells
2
Transwell Assay for PASMC Migration
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Migration of PASMCs was determined by Transwell assay, as described previously [18 ]. Shortly, 50,000 cells were seeded on the top of each polycarbonate filter with 8-μm pores (Corning) in 0.1 ml of basal medium, and then stimulated by PDGF (20 ng/ml) with or without Phps-1 (20 μM) for 12 h. Following exposure, the cells were fixed in 4% paraformaldehyde for 30 min and stained with 5% crystal violet (Beytime, China) for 30 min. Unmigrated cells were then scraped off the top of the filter. For each filter, at least five randomly chosen fields were imaged to obtain a total cell count.
3
Transwell Assay for Cell Migration
4
Quantifying Schwann Cell Migration
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Schwann cells transfected with let-7 mimic, let-7 inhibitor, or the corresponding controls were resuspended in DMEM (Corning) and seeded onto the upper chamber of a 6.5-mm transwell with 8-μm pores (Corning) at a volume of 100 μL and a density of 3 × 105 cells/mL. The bottom chamber of the transwell was filled with 500 μL cell complete culture medium as described above. After 24 hours of incubation, Schwann cells remaining on the upper surface of the upper chamber were cleaned with a cotton swab, and Schwann cells that migrated to the bottom surface were stained with 0.1% cresyl violet (Cat# C8470, Solarbio, Beijing, China). Images were captured with a Leica DMI3000 B (Leica Microsystems). Migrated cells were dissolved in cresyl violet with 33% acetic acid (Cat# 64-19-7, Xilong Scientific, Guangzhou, Guangdong Province, China) and the absorbance of cresyl violet staining was measured using a Synergy™ 2 Multi-Mode Microplate Reader (BioTek, Burlington, VT, USA).
5
Metformin Inhibits TGF-β1-induced Cell Migration
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NMuMG cells were treated with TGF-β1 for 36 h, 2 × 105 cells in the 0.2 % FBS medium were plated into the upper chamber with 8-μm pores (Corning; Life Sciences, MA, USA), and DMEM containing 10 % FBS was placed in the lower chamber. Cells were treated with metformin (10 mM) in the upper chamber and TGF-β1 (5 ng/ml) in the lower chamber. After 24 h, the filters were fixed in methanol for 10 min and stained with 0.1 % crystal violet (Solabri, Beijing) for 12 min. Randomly selected fields were photographed under a microscope, and stained cells were statistically analyzed.
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