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

被引:55
作者
Bai, Yan [1 ]
Krishnamoorthy, Nandini [2 ]
Patel, Kruti R. [1 ]
Rosas, Ivan [2 ]
Sanderson, Michael J. [3 ]
Ai, Xingbin [1 ,2 ]
机构
[1] Boston Univ, Sch Med, Pulm Allergy Sleep & Crit Care Med, Boston, MA 02215 USA
[2] Brigham & Womens Hosp, Pulm & Crit Care Med, 75 Francis St, Boston, MA 02115 USA
[3] Univ Massachusetts, Sch Med, Dept Microbiol & Physiol Syst, Worcester, MA 01655 USA
基金
美国国家卫生研究院;
关键词
human precision-cut lung slices; cryopreservation; Ca2+ oscillations; bitter-taste receptor; biobank; AIRWAY SMOOTH-MUSCLE; CA2+ OSCILLATIONS; CALCIUM OSCILLATIONS; CELLS; CONTRACTION; RELAXATION; SENSITIVITY; SENSITIZATION; PRINCIPLES; KINASE;
D O I
10.1165/rcmb.2015-0290MA
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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 Ca2+-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 Ca2+ 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.
引用
收藏
页码:656 / 663
页数:8
相关论文
共 46 条
[1]   Diverse macrophage populations mediate acute lung inflammation and resolution [J].
Aggarwal, Neil R. ;
King, Landon S. ;
D'Alessio, Franco R. .
AMERICAN JOURNAL OF PHYSIOLOGY-LUNG CELLULAR AND MOLECULAR PHYSIOLOGY, 2014, 306 (08) :L709-L725
[2]   TAS2R activation promotes airway smooth muscle relaxation despite β2-adrenergic receptor tachyphylaxis [J].
An, Steven S. ;
Wang, Wayne C. H. ;
Koziol-White, Cynthia J. ;
Ahn, Kwangmi ;
Lee, Danielle Y. ;
Kurten, Richard C. ;
Panettieri, Reynold A., Jr. ;
Liggett, Stephen B. .
AMERICAN JOURNAL OF PHYSIOLOGY-LUNG CELLULAR AND MOLECULAR PHYSIOLOGY, 2012, 303 (04) :L304-L311
[3]   Agonist-induced Ca2+ Sensitization in Smooth Muscle REDUNDANCY OF RHO GUANINE NUCLEOTIDE EXCHANGE FACTORS (RhoGEFs) AND RESPONSE KINETICS, A CAGED COMPOUND STUDY [J].
Artamonov, Mykhaylo V. ;
Momotani, Ko ;
Stevenson, Andra ;
Trentham, David R. ;
Derewenda, Urszula ;
Derewenda, Zygmunt S. ;
Read, Paul W. ;
Gutkind, J. Silvio ;
Somlyo, Avril V. .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2013, 288 (47) :34030-34040
[4]  
Baatz JE, 2014, IN VIVO, V28, P411
[5]   Airway smooth muscle relaxation results from a reduction in the frequency of Ca2+ oscillations induced by a cAMP-mediated inhibition of the IP3 receptor [J].
Bai, Y ;
Sanderson, MJ .
RESPIRATORY RESEARCH, 2006, 7 (1)
[6]   Modulation of the Ca2+ sensitivity of airway smooth muscle cells in murine lung slices [J].
Bai, Yan ;
Sanderson, Michael J. .
AMERICAN JOURNAL OF PHYSIOLOGY-LUNG CELLULAR AND MOLECULAR PHYSIOLOGY, 2006, 291 (02) :L208-L221
[7]   The cryopreservation of composite tissues Principles and recent advancement on cryopreservation of different type of tissues [J].
Bakhach, Joseph .
ORGANOGENESIS, 2009, 5 (03) :119-126
[8]   Acetylcholine-induced calcium signaling and contraction of airway smooth muscle cells in lung slices [J].
Bergner, A ;
Sanderson, MJ .
JOURNAL OF GENERAL PHYSIOLOGY, 2002, 119 (02) :187-198
[9]   Smooth muscle cell calcium activation mechanisms [J].
Berridge, Michael J. .
JOURNAL OF PHYSIOLOGY-LONDON, 2008, 586 (21) :5047-5061
[10]   Interleukin-13 Augments Bronchial Smooth Muscle Contractility with an Up-Regulation of RhoA Protein [J].
Chiba, Yoshihiko ;
Nakazawa, Shuji ;
Todoroki, Michiko ;
Shinozaki, Koji ;
Sakai, Hiroyasu ;
Misawa, Miwa .
AMERICAN JOURNAL OF RESPIRATORY CELL AND MOLECULAR BIOLOGY, 2009, 40 (02) :159-167