Invasion assays were performed using a HTS Transwell-24 system (Corning, Inc.). The Transwell inserts were coated with Matrigel (BD Biosciences) for 20 min at room temperature. At 48 h post-transfection, cells were collected after trypsin digestion. Subsequently, cells (1x105) in 200 µl serum-free medium were plated into the upper chamber. Medium (300 µl) supplemented with 10% FBS was plated into the lower chamber. After incubation for 24 h at 37˚C, non-invading cells on the upper surface of the inserts were gently removed with a cotton swab. Invading cells were fixed with 50% methanol for 10 min at room temperature and stained with 0.01% crystal violet for 15 min at room temperature. Invasive cells were imaged using an Olympus IX71 inverted microscope (Olympus Corporation; magnification, x400) and analysed using ImageJ1. 50i software (National Institutes of Health).
Hts transwell 24 system
Manufactured by Corning
Sourced in United States
The HTS Transwell-24 system is a high-throughput cell culture platform designed for advanced in vitro studies. It features a 24-well format with permeable membrane supports, enabling parallel analysis of multiple cell models and drug transport assays.
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4 protocols using hts transwell 24 system
1
Matrigel-Based Cell Invasion Assay
2
Transwell Assays for Cell Migration and Invasion
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Transwell assays were conducted to evaluate the cell migration and invasion using the HTS transwell-24 system (Corning, NY, USA), which is an array of 24 individual Boyden chambers with 8-μm pore size transwell membranes. Cell migration assays were performed as previously described [45 (link)]. For invasion assays, the upper chambers were coated with matrigel. Cells on the lower surface were fixed, stained and counted at 8–12 h after seeding.
3
Transwell Assay for Cell Invasion
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Transwell assays were performed to assess cell invasion using HTS Transwell‐24 system (Corning Inc., Corning, NY, USA). Cells were cultured in serum‐free RPMI 1640 medium for 24 h, followed by the addition of 100 µL of 5 × 104 cells into the upper chamber, whereas the lower chamber was filled with RPMI 1640 medium supplemented with 10% fetal bovine serum as a chemoattractant. Cells from the serum‐free upper chamber have a tendency to pass through the membrane because of the presence of serum‐containing medium in the lower compartment, which allowed the estimation of cell migration and invasion ability. After incubation at 37 °C for 24 h, cells in the lower compartment that had passed through the membrane were stained with Giemsa, fixed with 4% paraformaldehyde on the slides and then counted under a light microscope.
4
Transwell Assay for Cell Invasion
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Transwell assays were performed to assess cell invasion using the HTS Transwell-24 system (Corning Inc., Corning, NY, USA). Cells were cultured in serum-free RPMI-1640 medium for 24 h. Then, 100 µL of 5 × 10 4 cells were seeded onto the upper chamber, and the lower chamber was filled with 600 µL of RPMI-1640 medium supplemented with 10% FBS as a chemoattractant. The cells in the upper chamber (without serum) have a tendency to pass through the membrane to the culture medium containing 10% FBS in the lower compartment, allowing an estimation of migratory and invasive abilities. After incubation at 37°C for 24 h, the cells that passed through the membrane and were in the lower compartment were stained with 0.2% crystal violet, fixed with 30% glycerin on the slides, and then counted under a light microscope.
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