Quantification of Mouse Tracheal Epithelial Cell Differentiation with Cilia Defects

Author

• Olivia V. Allen, University of South DakotaFollow

Author ORCID Identifier

https://orcid.org/0009-0006-3944-7944

Date of Award

Spring 4-23-2026

Document Type

Honors Thesis

Department/Major

Basic Biomedical Science

Additional Department

Health Science

First Advisor

Musheera Abdellatif

Second Advisor

Lance Lee

Third Advisor

Mark Dixon

Keywords

Primary Ciliary Dyskinesia, Motile Cilia, Mouse Tracheal Epithelial Cells

Subject Categories

Cell and Developmental Biology | Cell Biology | Developmental Biology | Diseases | Genetics and Genomics | Life Sciences | Medicine and Health Sciences | Molecular Genetics | Respiratory Tract Diseases

Abstract

Primary Ciliary Dyskinesia (PCD) is an autosomal recessive disorder that affects 1 in every 7,500 live births. PCD results from motile cilia defects, impairing mucociliary clearance and causing chronic respiratory infections. Our lab studies include three PCD mouse lines — bgh (Spef2), nm1054 (Cfap221), and Cfap54^gt/gt (Cfap54). Prior transcriptomic data led us to hypothesize that PCD mutant mouse tracheal epithelial cells (MTECs) exhibit reduced differentiation into ciliated cells. MTECs from the trachea of the Wild Type (WT) and the PCD mutant mouse lines were cultured at an Air Liquid Interface (ALI) to start the differentiation into ciliated cells. On ALI day 5, cells were fixed and stained with DEUP1 by Immunofluorescence to count deuterosomal cells. On ALI day 13, MTECs were fixed and stained with acetylated tubulin to count the ciliated cells. Cells were imaged on the Nikon SoRa Super Resolution Confocal microscope. NIS elements was used to manually quantify the deuterosomal and ciliated cells from the images obtained from the SoRa. MTECs were analyzed for Ciliary Beat Frequency (CBF) on the Olympus IX71 microscope using the Sisson Ammons Video Analysis (SAVA) software. On ALI day 5, PCD cultures showed higher deuterosomal cell counts than WT; by day 13, ciliated cell counts were reduced in PCD cultures compared to WT. Quantification of the beating cilia area on ALI day 13 using the IX71 microscope and SAVA analysis software showed PCD mutants had a significantly lower beating area compared to the WT. Findings suggest that differentiation of ciliated cells may be down-regulated because of PCD pathogenesis.

Recommended Citation

Allen, Olivia V., "Quantification of Mouse Tracheal Epithelial Cell Differentiation with Cilia Defects" (2026). Honors Thesis. 432.
https://red.library.usd.edu/honors-thesis/432