An in vitro invasion assay was carried out to examine the invasion of cervical cancer cells, as previously described [21 (link)]. Briefly, 24-well Transwell units with 8 µm polycarbonate nucleopore filters (Corning, NY, USA) were coated with 0.1 mL 0.8 mg/mL Engelbreth Holm-Swarm sarcoma tumor extract (EHS Matrigel) at room temperature for 1 h to form a genuinely reconstituted basement membrane. Cervical cancer cells (5 × 104 cells) were placed in the upper compartment and 500 µL DMEM culture medium containing 10% fetal calf serum was added to the lower compartment. The Transwell plates were incubated at 37 °C for 36 h in a humidified atmosphere with 5% CO2 and stained with 10% crystal violet. Invading cells were defined as cells that had degraded the Matrigel and moved into the lower surface of the membrane. The non-invading cells retained on the upper surface of the membrane were removed by a cotton swab.
Polycarbonate nucleopore filters
Manufactured by Corning
Sourced in United States
Polycarbonate nucleopore filters are laboratory equipment used for filtration processes. They are designed with a uniform pore structure that allows for the precise separation and isolation of particles, cells, or molecules based on size.
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Lab products found in correlation
2 protocols using polycarbonate nucleopore filters
1
Cervical Cancer Invasion Assay
2
Transwell Assays for Cell Migration and Invasion
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The migration and invasion of CCA were examined by transwell assay. After 24 hrs of transfection, cells (5×104 cells/well) were detached with serum free RPMI-1640 and loaded in the upper section of a 24-well transwell unit with 8 μm polycarbonate nucleopore filters (Corning, NY, USA), while medium with 10% FBS was added to the lower chamber. The transwell unit was incubated for 24 hrs before the cells on the lower surface of the membrane were fixed and stained with crystal violet. The experiment was performed in triplicate. For the invasion assay, 40 μL Matrigel (BD Biosciences, San Jose, CA, USA) was coated in the top filter of the transwell unit and placed in an incubator at 37 °C for 4 h to form a reconstructed basement membrane. The methods used were identical to those applied to the migration assay.
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