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.
The nuclear receptors pregnane X receptor (PXR) and constitutive androstane receptor (CAR) are closely related transcription factors that regulate the expression of phase I (cytochrome P450s), phase II metabolizing enzymes and transporter genes in response to xenobiotics, including prescription drugs.READ MORE
We utilized paired synthetic crRNAs coupled with our synthetic tracrRNA in cells transduced with lentiviral Cas9 to perform a functional knockout on hsa-miR-221. This three-part system (crRNA, tracrRNA and Cas9) has demonstrated efficient gene editing when used with only one guide RNA, but the goal was to use two crRNAs to remove the entire stem-loop.READ MORE
2nd International Conference on Pharmaceutical Research & Innovations in Pharma Industry
May 30 - May 31, 2019