title: Porcine nasal epithelial cultures for studies of cystic fibrosis sinusitis.
abstract: BACKGROUND
  Transgenic cystic fibrosis (CF) murine models do not develop spontaneous lung or sinus disease, 2 major causes of morbidity in human CF patients. Because of these limitations, transgenic cystic fibrosis transmembrane conductance regulator (CFTR)(-/-) pigs have been developed and are currently being characterized. These CF animal models have phenotypes closely resembling that of human CF subjects. The objectives of the current study were to develop primary porcine nasal epithelial (PNE) cultures and evaluate their usefulness as a means to investigate sinonasal transepithelial transport and CFTR function.
  
  
  METHODS
  PNE derived from the septum or turbinates of CFTR(+/+) and CFTR(-/-) pigs were cultured at an air-liquid interface to confluence and full differentiation. Epithelial monolayers were mounted in Ussing chambers to investigate pharmacologic manipulation of ion transport. Ciliary beat frequency (CBF) and scanning electron microscopy of monolayers were used to indicate degree of ciliation and cell differentiation.
  
  
  RESULTS
  Stimulation of CFTR-mediated anion transport (ΔIsc in μA/cm(2) ) was significantly greater in epithelia derived from the septum when compared to turbinates (33.04 ± 1.17 vs 18.9 ± 0.73; p < 0.05). Cyclic adenosine monophosphate (cAMP)-activated Cl(-) secretion was absent in CFTR(-/-) and present in CFTR(+/+) epithelia. Calcium-activated Cl(-) (CaCC) secretion was increased in CF; however, overall Cl(-) transport through CaCCs was very low. Degree of ciliation (90%) and CBF were similar between groups.
  
  
  CONCLUSIONS
  Septal PNE exhibit a robust ion transport phenotype and indicate CFTR(-/-) sinus disease could be attributable to diminished alternative pathways for Cl(-) transport. Overall, PNE have similarities to human respiratory epithelia not demonstrated in murine cells and represent useful in vitro models for studying CF sinus disease.
authors.affiliation: Department of Surgery, Division of Otolaryngology, University of Alabama at Birmingham, Birmingham, AL.
authors.forename: Nichole
authors.initials: N
authors.lastname: Dean
authors.forename: Neel K
authors.initials: NK
authors.lastname: Ranganath
authors.forename: Brandon
authors.initials: B
authors.lastname: Jones
authors.forename: Shaoyan
authors.initials: S
authors.lastname: Zhang
authors.forename: Daniel
authors.initials: D
authors.lastname: Skinner
authors.forename: Steven M
authors.initials: SM
authors.lastname: Rowe
authors.forename: Eric J
authors.initials: EJ
authors.lastname: Sorscher
authors.forename: Bradford A
authors.initials: BA
authors.lastname: Woodworth
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D002712:Chlorides / Q000378:metabolism; D003550:Cystic Fibrosis / Q000150:complications / Q000235:genetics / Q000276:immunology*; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000235:genetics; D004195:Disease Models, Animal; D006801:Humans; D017136:Ion Transport / Q000235:genetics; D051379:Mice; D018181:Mice, Inbred CFTR; D009297:Nasal Mucosa / Q000166:cytology*; D012852:Sinusitis / Q000150:complications / Q000235:genetics / Q000276:immunology*; D013552:Swine; D046509:Tissue Culture Techniques / Q000379:methods*
chemical_list: D002712:Chlorides; D019005:Cystic Fibrosis Transmembrane Conductance Regulator
keywords: CFTR; chloride secretion; mucociliary clearance; porcine nasal culture; sinusitis; transepithelial ion transport
references.citation: Am J Respir Cell Mol Biol. 2007 Jan;36(1):1-7
references.pmid: 16888286
references.citation: J Clin Invest. 2010 Sep;120(9):3149-60
references.pmid: 20739752
references.citation: J Clin Invest. 2008 Apr;118(4):1571-7
references.pmid: 18324337
references.citation: PLoS One. 2013 Nov 25;8(11):e81589
references.pmid: 24282612
references.citation: Am J Respir Cell Mol Biol. 2014 May;50(5):953-62
references.pmid: 24303840
references.citation: Proc Natl Acad Sci U S A. 1994 Jan 18;91(2):479-83
references.pmid: 7507247
references.citation: N Engl J Med. 2005 May 12;352(19):1992-2001
references.pmid: 15888700
references.citation: Biotechniques. 2007 Aug;43(2):195-6, 198, 200 passim
references.pmid: 17824387
references.citation: Science. 1992 Aug 21;257(5073):1083-8
references.pmid: 1380723
references.citation: Laryngoscope. 2011 Jun;121(6):1313-9
references.pmid: 21480283
references.citation: Mol Ther. 2013 May;21(5):947-53
references.pmid: 23511247
references.citation: J Microsc. 2003 Aug;211(Pt 2):103-11
references.pmid: 12887704
references.citation: Methods Mol Biol. 2002;188:115-37
references.pmid: 11987537
references.citation: Am J Rhinol. 2008 Jan-Feb;22(1):7-12
references.pmid: 18284852
references.citation: Cell. 2010 Dec 10;143(6):911-23
references.pmid: 21145458
references.citation: Am J Rhinol Allergy. 2011 Sep-Oct;25(5):307-12
references.pmid: 22186243
references.citation: Science. 2008 Oct 24;322(5901):590-4
references.pmid: 18772398
references.citation: Am J Physiol Lung Cell Mol Physiol. 2008 Aug;295(2):L240-63
references.pmid: 18487356
references.citation: Biosci Rep. 1995 Dec;15(6):531-41
references.pmid: 9156582
references.citation: Am J Rhinol. 2008 May-Jun;22(3):235-8
references.pmid: 18588754
references.citation: Am J Rhinol. 2007 Sep-Oct;21(5):533-7
references.pmid: 17999784
references.citation: Otolaryngol Head Neck Surg. 2010 Sep;143(3):397-404
references.pmid: 20723778
references.citation: Am J Rhinol Allergy. 2009 Mar-Apr;23(2):149-52
references.pmid: 19401039
references.citation: JAMA Otolaryngol Head Neck Surg. 2013 Aug 1;139(8):822-7
references.pmid: 23949358
references.citation: Laryngoscope. 2012 Sep;122(9):1898-905
references.pmid: 22711071
references.citation: Science. 2008 Sep 26;321(5897):1837-41
references.pmid: 18818360
references.citation: Expert Rev Mol Med. 2001 Mar 12;2001:1-27
references.pmid: 14987374
references.citation: Laryngoscope. 2011 Sep;121(9):1929-34
references.pmid: 22024847
references.citation: Biol Reprod. 2011 Sep;85(3):442-51
references.pmid: 21593481