For in vitro invasion assay, cancer cells (1.5 × 105 cells in 200 µL) were suspended in the upper half of a PET membrane transwell insert chamber (BD Biosciences), which was coated with Matrigel (1 mg/mL; BD Biosciences), on a 24-well plate. Medium supplemented with 10% FBS was added as a chemoattractant to the lower half. After incubation at 37 °C for 24 hr, cancer cells that passed through the insert were fixed with 3.7% formalin (Sigma-Aldrich) and stained with 0.1% crystal violet (Sigma-Aldrich).
Pet membrane transwell insert chamber
Manufactured by BD
The PET membrane transwell insert chamber is a lab equipment component used for cell culture and migration assays. It consists of a cell culture insert with a porous polyethylene terephthalate (PET) membrane that allows for the separation and study of cellular interactions between different cell types or conditions.
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4 protocols using pet membrane transwell insert chamber
1
In Vitro Cancer Cell Invasion Assay
2
Cancer Cell Migration and Invasion Assays
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For in vitro migration assay, cancer cells (70μL; concentration: 7×105 cells/mL) were added to Culture-Insert well (ibidi) and cultured for 24 hr. After removal of Culture-Insert, cancer cells were cultured for 20 hr. The migration distance of cancer cells was recorded and measured using ImageJ.
For in vitro invasion assay, cancer cells (1.5×105 cells in 200 μL) were suspended in DMEM medium and added to the upper half of a PET membrane transwell insert chamber (BD Biosciences), which was coated with Matrigel (1 mg/mL; BD Biosciences) on a 24-well plate. DMEM medium supplemented with 10% FBS was added as a chemoattractant to the lower half. After incubation at 37°C for 24 hr, cancer cells that passed through the insert were fixed with 3.7% formalin (Sigma-Aldrich) and stained with 0.1% crystal violet (Sigma-Aldrich).
For in vitro invasion assay, cancer cells (1.5×105 cells in 200 μL) were suspended in DMEM medium and added to the upper half of a PET membrane transwell insert chamber (BD Biosciences), which was coated with Matrigel (1 mg/mL; BD Biosciences) on a 24-well plate. DMEM medium supplemented with 10% FBS was added as a chemoattractant to the lower half. After incubation at 37°C for 24 hr, cancer cells that passed through the insert were fixed with 3.7% formalin (Sigma-Aldrich) and stained with 0.1% crystal violet (Sigma-Aldrich).
3
In Vitro Cancer Cell Invasion Assay
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For in vitro invasion assay, cancer cells (1.5 × 105 cells in 200 μL) were suspended in the upper half of a PET membrane transwell insert chamber (BD Biosciences), which was coated with Matrigel (1 mg/mL; BD Biosciences), on a 24-well plate. Medium without FBS supplement was added into the upper chamber, whereas medium with 10% FBS supplement was added into the lower chamber. After incubation at 37 °C for 24 h, cancer cells that passed through the insert were fixed with 3.7% formalin (Sigma-Aldrich) and stained with 0.1% crystal violet (Sigma-Aldrich).
4
Tumor Cell Migration and Invasion Assay
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The wound healing assay was used to study the migration ability of tumor cell in vitro. Tumor cells (70 µL; concentration: 3 × 105 cells/mL) were added to Culture‐Insert well (ibidi) and cultured for 24 h. A “wound gap” was created by removal of Culture‐Insert, and the “healing” of this gap by cell migration was recorded per 4 h until 24 h. The migration area of tumor cells was measured using Image J software. The Matrigel invasion assay was used to determine the invasion ability of tumor cell in vitro. Tumor cells (2.5 × 105 cells in 200 µL) were suspended in DMEM medium and added to the upper half of a PET membrane transwell insert chamber (BD Biosciences), which was coated with Matrigel (1 mg/mL; BD Biosciences) on a 24‐well plate. DMEM medium supplemented with 10% FBS was added as a chemoattractant to the lower half. After incubation at 37°C for 24 h, tumor cells that passed through the insert were fixed with 3% formalin (Sigma‐Aldrich) and stained with 0.2% crystal violet (Sigma‐Aldrich).
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