cB-cell clones were cultured in a Millicell culture insert.
As shown in the top panel, non-cystic fibrosis HBE develop a liquid meniscus that surrounds the culture insert.
The day before starting the co-cultures, cells were plated at a density of 1 × 104 cells/cm2 in each well of a 12-well Transwell culture insert.
After another treatment with Gap26 for 72 h, the cells on the culture insert were fixed with 4%% formaldehyde in PBS for subsequent immunocytochemistry.
Three days before starting the co-cultures, BMSCs were plated at a density of 1 × 104 cells/cm2 in each well of a 12-well Transwell culture insert.
On the day before the AECs were prepared, rat BMSCs were plated at a density of 2 × 105 cells/cm2 on the 12-mm Transwell culture insert.
Image depicts AECs on the culture insert of a Transwell on day 7, rotating about the y-axis to demonstrate the three-dimensional alveolar-like structure.
The second step consists of culturing the cells with their CM, after recovery from the beads, on a tissue culture insert with a porous membrane.
To resolve these problems, a novel multiple-funnel cell culture insert was designed for size controlling, easy harvesting, and scale-up production of cell spheres.
Following a brief centrifugation, the cells with their CM were recovered, resuspended in medium containing 10% or 20% FBS and seeded onto a tissue culture insert.
Membranes were detached from the culture insert and fixed in absolute ethanol and subsequently in acetone.
