Research Information
A number of experimental techniques that involve measurements in engineered CF cell lines (available through the Center’s Gene Expression Core) and tissues of CF animal models (available through the Center’s CFTR Rat Models Core and Animal and Preclinical Models Core) are essential for advancing CF science. Established in 1994, the Center’s Functional Assay Core provides investigators with training, expertise and specialized equipment to aid in the advancement of basic and translational science studies in CF in these model systems. Specific capabilities of the Core include:
Core Function #1: Provide the ability to monitor changes in ion transport including CFTR activity
Monitoring ion transport in CF and control cells and tissues can be used to characterize the CF defect and to monitor the response to agents that alter ion transport pathways, including pharmacologic agents intended to restore CFTR activity, a mainstay of research at our CF Center and beyond. The Core can conduct multiple complimentary assays in this area, focusing on analyses in CF cell lines and animal tracheal tissues:
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Ussing chamber analyses
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Transepithelial chloride conductance (TECC) using robots that calculate changes in conductance or equivalent current in high-throughput formats
Core Function #2: Provide measurements of mucociliary transport and fluid flux
Micro-optical coherence tomography (μOCT), co-invented by our Center, enables quantitative, real-time, and simultaneous analyses of airway surface liquid, periciliary layer depth, ciliary beat frequency, mucociliary transport, and mucus viscosity from the surface of airway mucosa. The Functional Assay Core conducts μOCT imaging of freshly harvested trachea with or without further experimental exposures ex vivo. Complementary in vitro, in vivo (animal models), and in-human analyses are available through the Center’s Cell Model and Evaluation, Animal and Preclinical Models, and Clinical and Translational Cores, respectively.
Organoid swelling enables assessment of fluid transport in cells from individual donors or engineered lines, helping accomplish our goal of hastening an era of individualized therapeutics. With the recent addition of high-speed video microscopy and a high-throughput image analysis, the Core provides quantitation of organoid swelling derived from airway cells, intestinal organoids, and iPS-derived spheroids.
Core Function #3: Provide patch clamp and single-channel analyses to study changes in CFTR channel gating and regulation
A number of mutations harbor dysregulated CFTR gating, or require analyses at the level of the individual cell; thus single-cell analyses are instrumental in defining this type of CF defect and to assist with mechanistic characterization. Given the current focus on personalized medicine and analyses of all CFTR mutations, this type of service has become a critical component of this Core, especially in cases where monolayer studies or organoids are not appropriate. The Core conducts:
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Patch clamp unitary conductance tracings to monitor open channel probability and test potentiators (or correctors) on a wide spectrum of CF mutations. Mutagenesis evaluations can be coupled to uncover mechanisms
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Macropatch of cells or excised tissues (e.g., sweat gland, airway gland) or fluorescent dye-based halide efflux methods (individual dye-loaded cells) to evaluate CFTR activity in cells grown on coverslips (SPQ assays) or isolated from excised tissues
Contact Information
Core users are associated both within and beyond the CF Research Center; there is no limitation on who may access the Core. For additional information regarding resources and services, please contact:
