HUVECs (6×104) were seeded in the upper chamber of Transwell inserts in 24-well plates (Corning, USA). Conditioned medium (CM) from U251 cells transfected with FMOD siRNA (si-FMOD) or control siRNA (si-Con) was added to the lower chamber. After culturing for 18 h, the migratory cells were counted via using a microscope.
Transwell inserts in
Manufactured by Corning
Sourced in United States
Transwell inserts are a type of laboratory equipment used for cell culture and in vitro experiments. They consist of a permeable membrane that separates the upper and lower chambers of a cell culture system, allowing for the study of cell migration, barrier function, and other cellular processes.
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2 protocols using transwell inserts in
1
HUVECs Migration Assay with FMOD Knockdown
2
Organoid Co-culture with Hydroxychloroquine
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FACS-sorted mouse club cells (5 × 103) or AT2 cells (2 × 104) were co-cultured with MLg (2 × 105), mouse lung fibroblast cells (ATCC, Rockefeller, MD, USA),
in Matrigel/culture medium (1:1) respectively. The culture medium included Dulbecco’s modified Eagle Medium/F12 (Corning, Corning, NY, USA), 1% insulin/transferrin/selenium (Sigma), 10% FBS,
100 IU/ml penicillin, 100 µg/ml streptomycin, and 10 µM SB431542 (Sigma). In the treatment group, 10 µM HCQ was added to the culture
medium. Then, the mixture was transferred to the Transwell inserts in 24-well plates (Corning) containing the culture medium at the bottom. An incubator with a humidified 37°C with 5%
CO2 was used to maintain cultures, and the culture medium was changed every other day. The organoids were imaged using an inverted fluorescence microscope (Olympus, Tokyo,
Japan). The colony-forming efficiency (CFE) was calculated as the number of organoids (greater than 50 µm in diameter) in each insert as a percentage of seeded epithelial
cells.
in Matrigel/culture medium (1:1) respectively. The culture medium included Dulbecco’s modified Eagle Medium/F12 (Corning, Corning, NY, USA), 1% insulin/transferrin/selenium (Sigma), 10% FBS,
100 IU/ml penicillin, 100 µg/ml streptomycin, and 10 µM SB431542 (Sigma). In the treatment group, 10 µM HCQ was added to the culture
medium. Then, the mixture was transferred to the Transwell inserts in 24-well plates (Corning) containing the culture medium at the bottom. An incubator with a humidified 37°C with 5%
CO2 was used to maintain cultures, and the culture medium was changed every other day. The organoids were imaged using an inverted fluorescence microscope (Olympus, Tokyo,
Japan). The colony-forming efficiency (CFE) was calculated as the number of organoids (greater than 50 µm in diameter) in each insert as a percentage of seeded epithelial
cells.
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