The goal of the core is to provide primary cells and cultured airway epithelial cell preparations from human, mouse and other species for the study of epithelial cells, lung biology and lung diseases.
- Primary culture human airway epithelial cells from normal and diseased tissues
- Primary culture airway epithelial cells from genetically defined strains of mice
- Media for primary airway cell culture
- Training of investigators in culture methods
- Consultation for analysis of gene expression and cell manipulation
Core procedures and capabilities: The airway epithelial cell core provides investigators with primary culture preparations of mouse and human airway epithelial cells differentiated at air-liquid interface to provide a model of normal airways including ciliated and secretory cells. The core utilizes a system in collaboration with the Thoracic Surgery Division for the routine procurement of human tissues from lung transplantation donors and explanted lungs and surgical tissues. The core can also culture airway cells obtained by lung biopsies, brushings, as well as nasal and sinus biopsy, brush, scrape or polyps. The core also has experience with the culture of trachea and lung tissues from mice, rats and pigs. Gene expression analysis protocols and training in methods are available.
Human samples: Core users may utilized human samples obtained and prepared by the core. Normal human airway epithelial cells can be obtained as de-identified tissues and exempted from specific human research. IRB permission may be required for some tissues and studies. Explanted tissues from patients with advanced lung diseases undergoing lung transplant are also available cultured on a limited bases. These are available with specific arrangements with the core. Advance lung diseases include: Interstitial lung diseases (primary idiopathic pulmonary fibrosis), chronic obstructive lung diseases, cystic fibrosis, and bronchiolitis obliterans.
Non-human preparations: The core has special expertise in the culture and differentiation of mouse tracheal epithelial cells (mTEC). Culture and differentiation of wild type mice and those with specific genetic backgrounds can performed. Users must provide mice and make specific arrangements. The core also has experience with the culture of airway epithelial cells from rats and pigs. Cells from other species can be cultured with some advanced planning.
Other lung cell types: Assistance with the isolation of lung fibroblasts and other cell types may be obtained with special planning through the core.
Manipulation and genetic modification of primary cells: The core has extensive experience with the manipulation of airway cells using growth factors, cytokines and specific culture conditions, as well as genetic manipulation using recombinant viral vectors. Assistance with lentivirus- and adenovirus-mediated gene expression or RNAi is available.
Cell function and gene expression analysis: Protocols and assistance with routine collection for protein and RNA are available. Assistance with techniques for in situ analysis of gene expression methods (e.g., immunofluorescence) can be provided. Assistance with live imaging of cilia function and models of airway clearance are available using core instruments including videomicroscopy.
Protocols and training: Protocols for cell isolation, culture and differentiation are available. Core users can be instructed on all methods for culture, manipulation and evaluation of protein and gene expression. Lead-time of one month should be provided to allow for scheduling and the necessary period for cell growth.
The core is available to investigators atWashington University and other non-profit organizations. Priorities are given to members of the Asthma and Allergic Diseases Cooperative Research Centers, and the lung research community with established relationships to the Pulmonary and Critical Care Division.
Please contact the core for information.
- Division of Pulmonary and Critical Care Medicine, Department of Medicine
- The facility is partially supported by awards from the Children’s Discovery Institute of Washington University School of Medicine and Children’s Hospital and from the National Institutes of Health (NHLBI) through an Asthma and Allergic Diseases Cooperative Research Centers award.
- Two hepafilter biosafety cabinets for tissue culture
- Two 37O C, CO2 incubators
- Nikon Dissecting Microscope
- Nikon Inverted tissue culture microscope with Hoffman Phase filters camera and fluorescent capabilities
- Nikon Eclipse Ti inverted phase microscope with phase contrast objectives and high speed video capturing capabilities and customized software (Sisson-Ammons Video Analysis/SAVA) within a temperature controlled environmental chamber