Abstract
Poster: 157Effects of common culture media on ionocyte morphology and function
Journal of cystic fibrosis, Vol.25(Suppl 2), pp.79-79
09/2026
DOI: 10.1016/j.jcf.2026.07.188
Abstract
When culturing cells for physiological investigation, it is critical to understand how in vitro conditions affect cellular phenotype and function. For primary human airway epithelia cultured at the air-liquid interface, a relatively small number of media formulations are used. Pneumacult Air Liquid Interface media (pcALI), Ultroser-G-containing media (USG), and Ham’s F-12 supplemented with pcALI (ALI-F12) are used to differentiate and maintain epithelia on transwell culture inserts. However, a systematic comparison of how these media influence the morphology and function of ionocytes has not been reported.
Primary human airway epithelial cells were isolated from donor lung tissue, enzymatically dissociated, and basal cells were purified and expanded using PneumaCult Ex-Plus. Expanded basal cells were seeded onto transwell culture inserts and differentiated at the air-liquid interface using pcALI, USG, or ALI-F12. Relative ionocyte numbers were quantified by flow cytometry, functional outcomes were assessed by measuring apical-to-basolateral Cl– flow through CFTR, and morphological outcomes were assessed by analyzing immunofluorescence images.
Preliminary flow analysis indicated differences in ionocyte population, with epithelia comprising 0.35% (ALF-12), 0.22% (USG), and 0.22% (pcALI) ionocytes. These findings are supported by our imaging and electrophysiology data. In response to an apical-to-basolateral [Cl–] gradient, epithelia generated CFTRinh-172-sensitive It values of 0.90 ± 0.38 (pcALI), 5.55 ± 3.19 (USG), and 6.75 ± 2.50 (ALI-F12) µA cm–2. When ionocytes were labeled with barttin antibodies and imaged on a confocal microscope, transwells cultured in pcALI contained markedly fewer ionocytes. Each medium exhibited distinct ionocyte morphology, ranging from semi-spherical in pcALI media to highly branched in USG media. The ALI-F12 media contained a mixture of branched and spherical ionocytes.
These data suggest that different culture media can influence physiological outcomes and ionocyte morphology. Key components and the physiological explanation for altered ionocyte population and morphology remain under investigation and may further our understanding of how the extracellular environment shapes ionocyte function in health and in cystic fibrosis.
Details
- Title: Subtitle
- Poster: 157Effects of common culture media on ionocyte morphology and function
- Creators
- A. Attar - University of IowaA. Clemons - University of Iowa, Internal MedicineS. Wu - University of IowaV. Srivastava - University of Iowa, Internal MedicineG Romano Ibarra - Division of Pulmonary, Critical Care, and Occupational Medicine, University of Iowa, Iowa City, IA, USAI. Thornell - University of Iowa
- Resource Type
- Abstract
- Publication Details
- Journal of cystic fibrosis, Vol.25(Suppl 2), pp.79-79
- DOI
- 10.1016/j.jcf.2026.07.188
- ISSN
- 1569-1993
- Publisher
- Elsevier B.V
- Number of pages
- 1
- Language
- English
- Date published
- 09/2026
- Academic Unit
- Pulmonary, Critical Care, and Occupational Medicine; Internal Medicine
- Record Identifier
- 9985237507102771
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