Investigations in the Cellular and Molecular Biology of Human Airway Mucociliary Tissue

Author(s)

Vincent Manna, Rowan University

Date Approved

8-2021

Document Type

Dissertation

Degree Name

PhD in Cell and Molecular Biology

Department

Molecular Biology

College

Graduate School of Biomedical Sciences

Sponsor

New Jersey Health Foundation

First Advisor

Salvatore Caradonna, PhD

Committee Member 1

Renee Demarest, Phd

Committee Member 2

Dimitri Pestov, PhD

Committee Member 3

Susan Muller-Weeks, PhD

Committee Member 4

Sergei Borukhov, PhD

Subject(s)

Respiratory System, Respiratory Mucosa, Nasal Mucosa, Investigative Techniques, Vitamin D, Angiotensin-Converting Enzyme 2, Vaping, Humans

Disciplines

Cell Biology | Circulatory and Respiratory Physiology | Investigative Techniques | Laboratory and Basic Science Research | Life Sciences | Medical Cell Biology | Medical Molecular Biology | Medicine and Health Sciences | Molecular Biology | Other Analytical, Diagnostic and Therapeutic Techniques and Equipment | Physiological Processes | Research Methods in Life Sciences

Abstract

Our laboratory has integrated the use of a human-derived, in vitro model of airway mucociliary tissue. We isolate human nasal epithelial cells (HNECs) from the nasal mucociliary tissue of donors with a small brush and expand the airway progenitor cells in culture. The HNECs are then seeded onto semi-permeable transwell inserts. The inserts are in contact with the media in the lower chamber but don’t contain media in the upper chamber therefore the cells are exposed to the air while drawing nutrients from the media below, this is called the Air-Liquid Interface (ALI). HNECs cultured at the ALI initiate a differentiation program and the progenitor cells regenerate fully functioning airway mucociliary tissue that maintains the morphology and cell types of mucociliary tissue located in the lung, bronchus, and nasal passages. This model system allows for investigations in the regenerative ability of HNECs, and fully differentiated ALI-tissue can be maintained in culture for weeks at a time for long-term experiments in the homeostatic maintenance of mucociliary tissue. At the conclusion of experiments ALI tissue can be harvested for analyses including histology, immunohistochemistry, gene expression studies, western blotting, and other molecular techniques. We identified an opportunity to write a comprehensive methods paper containing all protocols necessary to implement the ALI model system and perform histological analyses. The complete method manuscript can be found in Chapter 1 of this dissertation. Upon implementation of the ALI model system our laboratory launched multiple lines of investigation into the biology of airway mucociliary tissue. Chapters 2, 3 and 4 are each unique research projects and all three utilized the ALI model system. Chapter 2 is an investigation into the roles of vitamin D (VD) and the vitamin D receptor (VDR) in ALI tissue. Chapter 3 is an examination of angiotensin converting enzyme 2 (ACE2) in ALI tissue. Chapter 4 combines the ALI model system with a novel vaporizing research device (VRD) created by the laboratory to investigate the impact that vape-pen vapor has on airway mucociliary tissue.

Recommended Citation

Manna, Vincent, "Investigations in the Cellular and Molecular Biology of Human Airway Mucociliary Tissue" (2021). Graduate School of Biomedical Sciences Theses and Dissertations. 33.
https://rdw.rowan.edu/gsbs_etd/33