Transwell system

Manufactured by Thermo Fisher Scientific

The Transwell system is a cell culture insert that allows for the separation of two compartments within a single well. It is designed to facilitate the study of various cellular processes, such as cell migration, permeability, and co-culture experiments.

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Lab products found in correlation

M csf, by Thermo Fisher Scientific (1 mentions) Anti cd3, by BD (1 mentions) Anti cd28, by BD (1 mentions) Celltracker green cmfda dye, by Thermo Fisher Scientific (1 mentions) Matrigel, by Corning (1 mentions) Eclipse te2000 u, by Nikon (1 mentions)

Close spelling variants of this product

NUNC® carrier plate transwell system

3 protocols using transwell system

Macrophage-Sca-1+ Cell Crosstalk Scaffold

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Bone marrow derived monocytes were obtained from the freshly isolated bone marrow of the femur and tibia of 2 weeks old SD rats. The monocytes were isolated using the differential adhesion method as previously reported [32 (link),33 (link)]. Macrophages were differentiated from bone marrow derived monocytes in the presence of macrophage colony stimulating factor (M-CSF, Peprotech) and then seeded on the PBS, Rapa, and 3-MA loaded scaffolds. The circular scaffold was composed of two parts, the lower PGS scaffold and upper PCL sheath. All of them were 2 mm thick and 15 mm in diameter. After 7 days of culture, cells adhered on the scaffolds were harvested for further analysis.
To investigate the crosstalk between macrophages seeded on drug-loaded scaffolds and PVAT-derived Sca-1+ cells, rat PVAT-derived Sca-1+ cells were cocultured with macrophages via the transwell system (pore size, 0.4 μm; Thermo Scientific). After coculturing for 7 days, Sca-1+ cells in the upper chamber were collected for further analysis. Each experiment was repeated in triplicate at least three times.

Chen W., Xiao W., Liu X., Yuan P., Zhang S., Wang Y, & Wu W. (2021). Pharmacological manipulation of macrophage autophagy effectively rejuvenates the regenerative potential of biodegrading vascular graft in aging body. Bioactive Materials, 11, 283-299.

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T cell activation and phenotyping

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Freshly isolated T cells were stimulated with 0.25 μg/mL of anti-CD3 (BD Biosciences) and 0.1 μg/mL of anti-CD28 (BD Biosciences) antibodies according to our previous reports [55 (link),56 (link)]. In some studies, recombinant human Gal-9 (GalPharma Co., Ltd, Japan) 1 μg/mL (approximately 31 μM) was added to T cells in the presence/absence of 100 μg/mL of an anti-CD44 blocking antibody (R&D Systems) or anti-CD137/4-1BB (Novus Biologicals). After 48 hours, T-cell activation and phenotype were measured using flow cytometry. In some studies, T-cell stimulation was performed using a trans-well system (Thermo Fisher Scientific) according to the manufacturer’s instructions.

Dunsmore G., Rosero E.P., Shahbaz S., Santer D.M., Jovel J., Lacy P., Houston S, & Elahi S. (2021). Neutrophils promote T-cell activation through the regulated release of CD44-bound Galectin-9 from the cell surface during HIV infection. PLoS Biology, 19(8), e3001387.

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Cell Migration and Invasion Assay

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Migration and invasion assays were performed using an 8 μm pore size Transwell system (Thermo Fischer Scientific). First, 2 × 10⁵ cells, were incubated with CellTracker™ Green CMFDA Dye (Invitrogen) for 30 min at 37 °C with following washing step with PBS. Cells resuspended in 200 ul of growth medium without FBS were added inside the insert fixed in the well with 600 ul medium with 10 % FBS. For invasion assays, the chamber was pre-coated with 1 mg/ml Matrigel (Corning). Cells were incubated for 3 h (migration) or 6 h (invasion) at 37 °C. Cells on the apical side of the chamber were gently scraped off using cotton swabs, filters were washed in PBS and pictures were taken under the microscope Nikon Eclipse TE 2000-U. Pictures analysis was done with the use of FIJI image processing package [19 (link)].

Przygodzka P., Sochacka E., Soboska K., Pacholczyk M., Papiewska-Pająk I., Przygodzki T., Płociński P., Ballet S., De Prins A, & Boncela J. (2021). Neuromedin U induces an invasive phenotype in CRC cells expressing the NMUR2 receptor. Journal of Experimental & Clinical Cancer Research : CR, 40, 283.

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