An HTS-Compatible Plate For Highly Miniaturized Cultures Of Primary Human Bronchial Epithelial Cells At Air-Liquid Interface
Poster Jun 29, 2015
Elizabeth Vu1, Eric Sorscher2, Robert Lowery1, Steven Hayes1
An obstacle to use of HBE-ALI models for high throughput screening (HTS) has been the severely limited supply of tissue samples from patients with unusual CFTR mutations. We have developed a plate for highly miniaturized, arrayed HBE-ALI cultures suitable for use in automated drug screening and mechanistic studies (μALI Plate). Cells cultured on the μALI Plate exhibit hallmarks of HBE biology including beating cilia and mucus production, but require less than one tenth the number of cells compared to conventional Transwell filters. Cells grown in this manner can be visualized directly from the apical surface by immunohistochemistry without the requirement to image through a filter support. Primary cells grown on the μALI Plate will be useful for testing therapeutic interventions directed towards patients with uncommon CFTR variants. Studies of ion channel function, cilia beat frequency and airway surface liquid depth are currently underway. Among other things, the μALI technology has the potential to facilitate personalized approaches to CF therapy, including drug development.
Basic fibroblast growth factor (bFGF) is widely used in vitro for the maintenance and stimulation of a variety of cells. However, use of native bFGF in cell biology is limited by the fact that bFGF rapidly degrades at physiological temperatures. We have addressed this problem with an engineered form of bFGF, named Heat Stable bFGF (HS bFGF), which is stable at 37 degrees Celsius.READ MORE