The invasive abilities of HA-VSMCs were measured using 24-well Transwell® chambers with 8-µm pore filters (Corning Inc). Cntl, EV and si-COL6A1 cells (5×104 cells) treated or untreated with 20 ng/ml PDGF-BB, diluted in Medium 231 supplemented with SMGS, were seeded into the Matrigel® GFR (BD Biosciences)-coated Transwell® upper chambers, with the coating process at 37°C for 30 min. The lower chambers were filled with Medium 231 supplemented with SMGS. Following 24 h incubation, the Transwell membranes were stained using 0.1% crystal violet for 30 min at 37°C. The number of invasive cells in random 5 fields was then calculated from images captured using Olympus DSX100 optical microscope (Olympus Corporation; magnification, ×200).
Transwell chambers with 8 m pore filters
Manufactured by Corning
Sourced in United States
Transwell® chambers with 8-µm pore filters are a laboratory tool used for cell culture and analysis. The chambers feature a semi-permeable membrane with 8-micrometer pores, allowing for the study of cell migration, transport, and interaction between different cell types or cell-environment interactions.
Automatically generated - may contain errors
2 protocols using transwell chambers with 8 m pore filters
1
Measuring VSMC Invasion Capacity
2
Transwell Migration and Invasion Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Transwell chambers with 8-µM pore filters (Corning Inc., USA) were used in the transwell migration and invasion assay to analyze the metastasis ability of BC cells.
In the transwell migration assay, after transfection for 24 h, BC cells (1×104 cells/100 µL serum-free medium) were seeded into the uncoated upper chambers. Fetal bovine serum (FBS, 10%) in medium was added to fill the lower chambers to act as the chemoattractant. After 24-h incubation, un-migrated BC cells in the upper surface were scraped using a cotton swab, and migrated BC cells were fixed using 4% paraformaldehyde (Sangon Biotech, China) and dyed using crystal violet (Sangon Biotech). The number of migrated BC cells in five random fields was counted and averaged.
In the transwell invasion assay, 40 µL BD Matrigel matrix (BD Biosciences, USA) diluted in cold serum-free medium of 1:8 was used to pre-coat the upper chambers to simulate the extracellular matrix before seeding BC cells (7×104 cells/100 µL serum-free medium; after transfection for 24 h), and the remaining procedures were similar with transwell migration assay.
In the transwell migration assay, after transfection for 24 h, BC cells (1×104 cells/100 µL serum-free medium) were seeded into the uncoated upper chambers. Fetal bovine serum (FBS, 10%) in medium was added to fill the lower chambers to act as the chemoattractant. After 24-h incubation, un-migrated BC cells in the upper surface were scraped using a cotton swab, and migrated BC cells were fixed using 4% paraformaldehyde (Sangon Biotech, China) and dyed using crystal violet (Sangon Biotech). The number of migrated BC cells in five random fields was counted and averaged.
In the transwell invasion assay, 40 µL BD Matrigel matrix (BD Biosciences, USA) diluted in cold serum-free medium of 1:8 was used to pre-coat the upper chambers to simulate the extracellular matrix before seeding BC cells (7×104 cells/100 µL serum-free medium; after transfection for 24 h), and the remaining procedures were similar with transwell migration assay.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!