Bai Yan, Krishnamoorthy Nandini, Patel Kruti R, Rosas Ivan, Sanderson Michael J, Ai Xingbin
1 Pulmonary, Allergy, Sleep and Critical Care Medicine, Boston University School of Medicine, Boston, Massachusetts.
2 Pulmonary and Critical Care Medicine, Brigham and Women's Hospital, Boston, Massachusetts; and.
Am J Respir Cell Mol Biol. 2016 May;54(5):656-63. 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.
人精密切割肺片(hPCLSs)为转化研究提供了一种独特的离体模型。然而,人肺组织有限且不可预测的可用性极大地阻碍了它们的应用。在此,我们证明hPCLSs的冷冻保存有助于储存活的人肺组织以供常规使用。我们的结果表明,冷冻保存对包括吞噬细胞和T淋巴细胞在内的免疫细胞的总体细胞活力和重要功能影响很小。此外,冷冻保存后,气道对特定激动剂和拮抗剂的收缩和舒张反应分别未发生变化。在亚细胞水平上,冷冻保存的hPCLSs维持了用于控制气道平滑肌细胞收缩性的钙(Ca2+)依赖性调节机制。为了举例说明冷冻保存的hPCLSs在平滑肌研究中的应用,我们提供证据表明苦味受体(TAS2R)激动剂通过阻断气道平滑肌细胞中的钙(Ca2+)振荡来舒张气道。总之,冷冻保存的hPCLSs的储存为肺生理学和疾病的转化研究提供了一种强大的生物测定方法。