Cryopreserved Human Precision-Cut Lung Slices as a Bioassay for Live Tissue Banking. A Viability Study of Bronchodilation with Bitter-Taste Receptor Agonists

• Published in: American journal of respiratory cell and molecular biology Vol. 54; no. 5; pp. 656 - 663
• Main Authors: Bai, Yan, Krishnamoorthy, Nandini, Patel, Kruti R, Rosas, Ivan, Sanderson, Michael J, Ai, Xingbin
• Format: Journal Article
• Language: English
• Published: United States American Thoracic Society 01.05.2016
• Subjects: Antibiotics; Biological Assay - methods; Bronchodilator Agents - pharmacology; Calcium Signaling - drug effects; Cell division; Cell Survival - drug effects; Cryopreservation; Experiments; Flow cytometry; Humans; Lung - cytology; Lungs; Lymphocytes; Lymphocytes - drug effects; Lymphocytes - metabolism; Major Technical Advances; Microscopy; Muscle Contraction - drug effects; Muscle Relaxation - drug effects; Myocytes, Smooth Muscle - drug effects; Myocytes, Smooth Muscle - metabolism; Phagocytes - drug effects; Phagocytes - metabolism; Physiology; Receptors, G-Protein-Coupled - agonists; Receptors, G-Protein-Coupled - metabolism; Smooth muscle; Software; Studies; Taste - drug effects; Tissue Banks; Oregon; Ca2+ oscillations; bitter-taste receptor; human precision-cut lung slices; biobank; cryopreservation
• ISSN: 1044-1549; 1535-4989
• DOI: 10.1165/rcmb.2015-0290MA

Human precision-cut lung slices (hPCLSs) provide a unique ex vivo model for translational research. However, the limited and unpredictable availability of human lung tissue greatly impedes their use. Here, we demonstrate that cryopreservation of hPCLSs facilitates banking of live human lung tissue for routine use. Our results show that cryopreservation had little effect on overall cell viability and vital functions of immune cells, including phagocytes and T lymphocytes. In addition, airway contraction and relaxation in response to specific agonists and antagonists, respectively, were unchanged after cryopreservation. At the subcellular level, cryopreserved hPCLSs maintained Ca(2+)-dependent regulatory mechanisms for the control of airway smooth muscle cell contractility. To exemplify the use of cryopreserved hPCLSs in smooth muscle research, we provide evidence that bitter-taste receptor (TAS2R) agonists relax airways by blocking Ca(2+) oscillations in airway smooth muscle cells. In conclusion, the banking of cryopreserved hPCLSs provides a robust bioassay for translational research of lung physiology and disease.