Polyester membrane transwell inserts (Corning Inc. Tewksbury, MA, 6.5 mm, 0.4 μm) containing monolayer or encapsulated MSCs (2.5 × 104, 5 × 104, or 1 × 105 cells/transwell) were added to host cultures in 24 well plates, and maintenance medium was exchanged for DMEM + 1% FBS, supplemented with 100 units/ml penicillin and 100 μg/ml streptomycin (“low serum media”) ±1 μg/ml LPS (Escherichia coli 055:B5, Sigma–Aldrich, St. Louis, MO).22 (link),23 (link) Nonstimulated and stimulated host cultures without MSC co-culture were used as controls. Cultures were returned to incubators at 37°C in 5% CO2 for 6, 12, 24, or 48 h, after which media supernatants were collected and cells were fixed.
Polyester membrane transwell inserts
Manufactured by Corning
Sourced in Morocco, United States
Polyester membrane transwell inserts are lab equipment used for cell culture applications. These inserts feature a porous polyester membrane that allows for the passage of cells, media, and other small molecules between the upper and lower chambers of the transwell system.
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Lab products found in correlation
3 protocols using polyester membrane transwell inserts
1
Transwell Immunomodulation Assay with MSCs
2
Corneal Epithelial Wound Healing Dynamics
3
Macrophage-Mesenchymal Stem Cell Interactions
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The RAW 264.7 macrophage cells (1 × 106 cells) were plated in 24 mm polyester membrane transwell inserts (0.4 μm pore size, Corning, USA) and allowed to attach for 24 h prior to treatment with lipopolysaccharide (LPS) Sigma-Aldrich, USA) for M1 phenotype induction. Macrophage cells were treated with 1 µg/mL LPS in DMEM medium supplemented with 10% FBS and 1% P/S for 24 h. The transwell inserts’ lower compartments were seeded with hTMSCs (1 × 106 cells), and placed with 2 mL hydrogel or 2 mL hydrogel with hTMSCs of the same cell number the following day. Co-culture with macrophage cells was kept for 24 h in DMEM medium supplemented with 10% FBS and 1% P/S. All cultures were kept at 37oC and 5% CO2. Macrophage cells were collected the next day and prepared for RNA extraction.
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