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Transwell migration system

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The Transwell Migration System is a laboratory equipment used to study cell migration and invasion. It consists of a multi-well insert with a porous membrane that allows cells to migrate from one chamber to another. The system facilitates the quantitative analysis of cell migration in response to various stimuli or experimental conditions.

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Lab products found in correlation

Matrigel invasion chamber, by BD (3 mentions) Ax10 microscope, by Zeiss (1 mentions) Matrigel, by BD (1 mentions) Polycarbonate filter, by Corning (1 mentions) Light microscopy, by Olympus (1 mentions)

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Transwell system (548 citations) Transwell migration assay system (2 citations)

6 protocols using transwell migration system

1

Invasion and Migration Assay Protocol

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Cell invasion was assayed using Matrigel Invasion Chambers (BD Biosciences). Briefly, cells were placed on the upper compartment of invasion chambers and treated with alcohol or MCP-1 in the presence or absence of CCR2 antagonist (CCR2-I). Culture medium containing 10% FBS was added into the lower compartment of invasion chambers and served as chemoattractants for the cells. Cells were maintained in the invasion chambers overnight. The invaded cells were fixed in 3.7% paraformaldehyde and stained with 0.5% crystal violet in 2% ethanol. Membranes were washed and the dye was eluted with 10% acetic acid. Absorbance was measured at 595 nm using a microtiterplate reader (Beckman coulter).
Cell migration was analyzed using a Transwell Migration System (Costar). Briefly, cells were plated into upper chambers (Transwells with 8.0 μm pore size) in serum free medium and treated with alcohol or MCP-1 in the presence or absence of CCR2 antagonist (CCR2-I). The lower compartment of the chamber contained regular medium containing 10% FBS. The chambers were cultured at 37°C in 5% CO2 for 12 hours. Migrated cells were fixed and stained with 0.5% crystal violet, followed by dye elution and absorbance measurement as described above.
Xu M., Wang S., Qi Y., Chen L., Frank J.A., Yang X.H., Zhang Z., Shi X, & Luo J. (2015). Role of MCP-1 in Alcohol-induced Aggressiveness of Colorectal Cancer Cells. Molecular carcinogenesis, 55(5), 1002-1011.
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2

Cell Invasion and Migration Assay

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Cell invasion was assayed using Matrigel Invasion Chambers (BD Biosciences). Cell migration was analyzed using a Transwell Migration System (Costar). Briefly, equal amount of cells were placed on the upper compartment of invasion chambers or Transwell chambers (8.0 μm pore size) which contained serum-free medium. Culture medium containing 10% FBS was added to the lower compartment of invasion/migration chambers to serve as chemoattractants. The chambers were cultured at 37°C in 5% CO2 in the presence or absence of ethanol (100 mg/dl) for 12 hours. The invaded/migrated cells were fixed in 4% paraformaldehyde and stained with 0.5% crystal violet in 2% ethanol. Membranes were washed and the dye was eluted with 10% acetic acid. Absorbance was measured at 595 nm using a microtiter plate reader (Beckman coulter).
Xu M., Wang S., Ren Z., Frank J.A., Yang X.H., Zhang Z., Ke Z.J., Shi X, & Luo J. (2015). Chronic ethanol exposure enhances the aggressiveness of breast cancer: the role of p38γ. Oncotarget, 7(3), 3489-3505.
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3

Chemotaxis Assay of PMN-MDSCs

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Chemotaxis was assayed using a Transwell migration system (5 μm; Costar). Because at least 95% of the splenic CD15+ mononuclear cells are CD15+ PMN-MDSCs, we isolated PMN-MDSCs using anti-CD15 magnetic beads. Following this isolation, the magnetically isolated cells were >98% PMN-MDSCs as described in Jordan et al. (21 (link)) 2.0 × 105 PMN-MDSCs were resuspended in serum free RPMI, seeded in the cell culture inserts, and placed in empty wells. Either RPMI, a solution containing 1 ng/ml IL-8 and RPMI, or 10% human serum in RPMI was added to corresponding wells. The culture was placed in a tissue culture incubator for 6 h at 37°C.
Cells were fixed in 10% neutral buffered formalin and stained with 0.2% crystal violet. Non-motile cells were scraped off the top of the chamber, and images of each filter were taken using a Zeiss AX10 microscope. Cells that had migrated to the bottom of the filter were counted on five representative areas of each 10X image using Image J (22 (link)). Each assay was repeated three times.
Tobin R.P., Jordan K.R., Kapoor P., Spongberg E., Davis D., Vorwald V.M., Couts K.L., Gao D., Smith D.E., Borgers J.S., Robinson S., Amato C., Gonzalez R., Lewis K.D., Robinson W.A., Borges V.F, & McCarter M.D. (2019). IL-6 and IL-8 Are Linked With Myeloid-Derived Suppressor Cell Accumulation and Correlate With Poor Clinical Outcomes in Melanoma Patients. Frontiers in Oncology, 9, 1223.
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4

Alcohol Exposure Effects on Cell Migration and Invasion

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Cell migration was analyzed using a Transwell Migration System (Costar). Cell invasion was assayed using Matrigel Invasion Chambers (BD Biosciences). Briefly, after alcohol exposure for 10 days, equal amount of cells were placed on the upper compartment of the Transwell chambers or invasion chambers in serum free medium. Culture medium containing 10 % FBS was added into the lower compartment of invasion/migration chambers and served as chemoattractants for the cells. The chambers were cultured at 37 °C in 5 % CO2 in the presence/absence of alcohol (100 mg/dl) for 12 h. Cells were fixed in 4 % paraformaldehyde and stained with 0.5 % crystal violet in 2 % ethanol. Membranes were washed and the cells that remained on the top of the invasion/transwell inserts were removed (non-migrated cells). The dye was eluted with 10 % acetic acid and the absorbance was measured at 595 nm using a microtiter platereader (Beckman coulter).
Xu M., Ren Z., Wang X., Comer A., Frank J.A., Ke Z.J., Huang Y., Zhang Z., Shi X., Wang S, & Luo J. (2016). ErbB2 and p38γ MAPK mediate alcohol-induced increase in breast cancer stem cells and metastasis. Molecular Cancer, 15, 52.
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5

Transwell Migration and Invasion Assay for HT29 Cells

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HT29 cells (6×104 cells/well) were seeded in the upper wells of a Transwell migration system attached with polycarbonate filters (Corning Incorporated, Corning, NY, USA) in RPMI-1640 supplemented with 0.1% FBS. The lower wells were filled with RPMI-1640 with 10% FBS. Following incubation for 24 h at 37°C, the non-migrating cells from the upper well were removed with cotton swabs. The cells that had migrated through the membranes were fixed with 70% cold ethanol at 4°C for 30 min, and stained by 0.1% crystal violet at 37°C for 30 min. Cells were counted in 5 separate fields of view and images were captured using light microscopy with ×200 magnification (Olympus Corporation, Tokyo, Japan) and cell migration rate was calculated.
The cell invasion assay was performed as aforementioned, with the exception of the application of chambers with Matrigel® (BD Biosciences, Franklin Lakes, NJ, USA), which was melted at 37°C for 30 min prior to usage.
Zhou T., Wu L., Wang Q., Jiang Z., Li Y., Ma N., Chen W., Hou Z., Gan W, & Chen S. (2018). MicroRNA-128 targeting RPN2 inhibits cell proliferation and migration through the Akt-p53-cyclin pathway in colorectal cancer cells. Oncology Letters, 16(6), 6940-6949.
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6

Wnt3-siRNA Inhibits MGC-803 Cell Migration

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The effect of Wnt3-siRNA on the migration of MGC-803 cells was analyzed using a transwell migration system (Corning Incorporated, Corning, NY, USA). After transfection with siRNAs for 48 hours, MGC-803 cells were digested and disrupted into single cell suspensions, equal numbers of cells (1,000 cells) were plated into the upper transwell chambers in 2% FBS-containing medium and 15% FBS-containing medium was added to the lower compartment of the chambers. The chambers with a pores sized of 8.0 μm were incubated at 37°C in 5% CO2 for 48 hours and cells remaining on the upper surface of the membrane were scratched by a cotton swab. The cells migrated through the pore and adhered to the lower surface were fixed with 4% paraformaldehyde for 20 minutes and stained with crystal violet (0.05%) for another 30 minutes before photographed with the microscope.
Wang H.S., Nie X., Wu R.B., Yuan H.W., Ma Y.H., Liu X.L., Zhang J.Y., Deng X.L., Na Q., Jin H.Y., Bian Y.C., Gao Y.M., Wang Y.D, & Chen W.D. (2016). Downregulation of human Wnt3 in gastric cancer suppresses cell proliferation and induces apoptosis. OncoTargets and therapy, 9, 3849-3860.
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