SUCCESSFUL ISOLATION AND CULTURE OF MULTIPOTENTIAL DISTAL AIRWAY STEM CELLS FROM COPD PATIENTS

Dale

Abstract

Chronic Obstructive Pulmonary Disease (COPD) presents a significant, incurable, worldwide health burden, identifying it as a prospective regenerative medicine target. Sourcing cells for lung therapies is challenging; progress has been made in culturing upper airway basal cells but information on the culture of distal cells remains scarce.

Distal lung from human donors was minced and digested. Cells were plated on type I collagen-coated flasks in cFAD medium with rock inhibitor (cFAD+)[1]. Cells were characterised by immunocytochemistry for vimentin, smooth muscle actin, pan-cytokeratin, TP63, cytokeratin-5, E-cadherin, Club cell secretory protein (CCSP), mucin-5AC and ß-IV-tubulin. Differentiated using air-liquid interface culture, with measurement of trans-epithelial electrical resistance (TEER), in a matrigel culture system, and seeded to porous collagen scaffolds.

Culture in cFAD+ resulted in predominantly epithelial cells expressing pan-cytokeratin and E-cadherin, TP63 and cytokeratin-5 suggesting a distal airway stem cell (DASC) identity. At air-liquid interface cells developed tight junctions with increased TEER (=350 O.cm2), increased levels of CCSP, stained positively for ß-IV-tubulin and mucin-5AC and had visible, motile cilia. Matrigel culture resulted in self-organising, mucus producing organoids, with visible motile cilia on the surface in addition to thin-walled, cavitated, alveolar-like organoids. The DASCs attached readily to collagen scaffolds, had excellent viability and exhibited proliferation on the scaffolds over a 14 day period.

We have successfully isolated and expanded epithelial progenitors from the distal lung tissue of COPD and healthy donors. Preliminary work demonstrates the DASC multipotential differentiation capacity in a variety of formats making them a valuable source for regenerative medicine therapeutic approaches.

Citation

Dale. (2022). SUCCESSFUL ISOLATION AND CULTURE OF MULTIPOTENTIAL DISTAL AIRWAY STEM CELLS FROM COPD PATIENTS. Tissue engineering. Part A, S451 - S452

• Acceptance Date: Apr 4, 2022
• Publication Date: Apr 4, 2022
• Journal: TISSUE ENGINEERING PART A
• Print ISSN: 1937-3341
• Publisher: Mary Ann Liebert
• Pages: S451 - S452
• Publisher URL: https://www.liebertpub.com/doi/10.1089/ten.tea.2022.29025.abstracts