#PAGE_PARAMS# #ADS_HEAD_SCRIPTS# #MICRODATA#

Equine bronchial fibroblasts enhance proliferation and differentiation of primary equine bronchial epithelial cells co-cultured under air-liquid interface


Autoři: Vanessa Abs aff001;  Jana Bonicelli aff001;  Johannes Kacza aff002;  Claudia Zizzadoro aff003;  Getu Abraham aff001
Působiště autorů: Institute of Pharmacology, Pharmacy and Toxicology, Faculty of Veterinary Medicine, University of Leipzig, An den Tierkliniken, Leipzig, Germany aff001;  Saxonian Incubator for Clinical Translation, University of Leipzig, Philipp-Rosenthal-Straße, Leipzig, Germany aff002;  Division of Veterinary Pharmacology and Toxicology, Department of Veterinary Medicine, University of Bari, SP 62 per Casamassima, km, Valenzano (BA), Italy aff003
Vyšlo v časopise: PLoS ONE 14(11)
Kategorie: Research Article
doi: https://doi.org/10.1371/journal.pone.0225025

Souhrn

Interaction between epithelial cells and fibroblasts play a key role in wound repair and remodelling in the asthmatic airway epithelium. We present the establishment of a co-culture model using primary equine bronchial epithelial cells (EBECs) and equine bronchial fibroblasts (EBFs). EBFs at passage between 4 and 8 were seeded on the bottom of 24-well plates and treated with mitomycin C at 80% confluency. Then, freshly isolated (P0) or passaged (P1) EBECs were seeded on the upper surface of membrane inserts that had been placed inside the EBF-containing well plates and grown first under liquid-liquid interface (LLI) then under air-liquid interface (ALI) conditions to induce epithelial differentiation. Morphological, structural and functional markers were monitored in co-cultured P0 and P1 EBEC monolayers by phase-contrast microscopy, scanning and transmission electron microscopy, hematoxylin-eosin, immunocytochemistry as well as by measuring the transepithelial electrical resistance (TEER) and transepithelial transport of selected drugs. After about 15–20 days of co-culture at ALI, P0 and P1 EBEC monolayers showed pseudo-stratified architecture, presence of ciliated cells, typically honeycomb-like pattern of tight junction protein 1 (TJP1) expression, and intact selective barrier functions. Interestingly, some notable differences were observed in the behaviour of co-cultured EBECs (adhesion to culture support, growth rate, differentiation rate) as compared to our previously described EBEC mono-culture system, suggesting that cross-talk between epithelial cells and fibroblasts actually takes place in our current co-culture setup through paracrine signalling. The EBEC-EBF co-culture model described herein will offer the opportunity to investigate epithelial-mesenchymal cell interactions and underlying disease mechanisms in the equine airways, thereby leading to a better understanding of their relevance to pathophysiology and treatment of equine and human asthma.

Klíčová slova:

Cell differentiation – Epithelial cells – Epithelium – Equines – Fibroblasts – Fluorescence imaging – Permeability – Tight junctions


Zdroje

1. Holgate ST, Lackie PM, Davies DE, Roche WR, Walls AF. The bronchial epithelium as a key regulator of airway inflammation and remodelling in asthma. Clin Exp Allergy 1999; 29 Suppl. 2: 90–95.

2. Xiao C, Puddicombe SM, Field S, Haywood J, Broughton-Head V, Puxeddu I, et al. Defective epithelial barrier function in asthma. J Allergy Clin Immunol. 2011; 128: 549–556. doi: 10.1016/j.jaci.2011.05.038 21752437

3. Humlicek AL, Manzel LJ, Chin CL, Shi L, Excoffon KJ, Winter MC, et al. Paracellular permeability restricts airway epithelial responses to selectively allow activation by mediators at the basolateral surface. J Immunol. 2007; 178: 6395–6403. doi: 10.4049/jimmunol.178.10.6395 17475869

4. Coyne CB, Vanhook MK, Gambling TM, Carson JL, Boucher RC, Johnson LG. Regulation of airway tight junctions by proinflammatory cytokines. Mol Biol Cell. 2002; 13: 3218–3234. doi: 10.1091/mbc.E02-03-0134 12221127

5. Haag S, Matthiesen S, Juergens UR, Racké K. Muscarinic receptors mediate stimulation of collagen synthesis in human lung fibroblasts. Eur Respir J. 2008; 32: 555–562. doi: 10.1183/09031936.00129307 18480105

6. Wenzel SE, Trudeau JB, Barnes S, Zhou X, Cundall M, Westcott JY, et al. TGF-beta and IL-13 synergistically increase eotaxin-1 production in human airway fibroblasts. J Immunol. 2002; 169: 4613–4619. doi: 10.4049/jimmunol.169.8.4613 12370400

7. Malavia NK, Mih JD, Raub CB, Dinh BT, George SC. IL-13 induces a bronchial epithelial phenotype that is profibrotic. Respir Res. 2008; 9: 27. doi: 10.1186/1465-9921-9-27 18348727

8. Skibinski G, Elborn JS, Ennis M. Bronchial epithelial cell growth regulation in fibroblast cocultures: the role of hepatocyte growth factor. Am J Physiol Lung Cell Mol Physiol. 2007; 293: L69–L76. doi: 10.1152/ajplung.00299.2006 17384084

9. Myerburg MM, Latoche JD, McKenna EE, Stabile LP, Siegfried JS, Feghali-Bostwick, CA, et al. Hepatocyte growth factor and other fibroblast secretions modulate the phenotype of human bronchial epithelial cells. Am J Physiol Lung Cell Mol Physiol. 2007; 292: L1352–L1360. doi: 10.1152/ajplung.00328.2006 17307814

10. Wiszniewski L, Jornot L, Dudez T, Pagano A, Rochat T, Lacroix JS, et al. Long-term cultures of polarized airway epithelial cells from patients with cystic fibrosis. Am J Respir Cell Mol Biol. 2006; 34: 39–48. doi: 10.1165/rcmb.2005-0161OC 16179582

11. Werner S, Smola H. Paracrine regulation of keratinocyte proliferation and differentiation. Trends Cell Biol. 2001; 11: 143–146. doi: 10.1016/s0962-8924(01)01955-9 11306276

12. Barton AK, Gehlen H. Pulmonary Remodeling in Equine Asthma: What Do We Know about Mediators of Inflammation in the Horse? Mediators Inflamm. 2016; 2016: 5693205. doi: 10.1155/2016/5693205 28053371

13. Bullone M, Lavoie JP. Asthma "of horses and men"—how can equine heaves help us better understand human asthma immunopathology and its functional consequences? Mol Immunol. 2015; 66: 97–105. doi: 10.1016/j.molimm.2014.12.005 25547716

14. Setlakwe EL, Lemos KR, Lavoie-Lamoureux A, Duguay JD, Lavoie JP. Airway collagen and elastic fiber content correlates with lung function in equine heaves. Am J Physiol Lung Cell Mol Physiol. 2014; 307: L252–L260. doi: 10.1152/ajplung.00019.2014 24879055

15. Leclere M, Lavoie-Lamoureux A, Joubert P, Relave F, Setlakwe EL, Beauchamp G, et al. Corticosteroids and antigen avoidance decrease airway smooth muscle mass in an equine asthma model. Am J Respir Cell Mol Biol. 2012; 47: 589–596. doi: 10.1165/rcmb.2011-0363OC 22721832

16. Abraham G, Zizzadoro C, Kacza J, Ellenberger C, Abs V, Franke J, et al. Growth and differentiation of primary and passaged equine bronchial epithelial cells under conventional and air-liquid-interface culture conditions. BMC Vet Res. 2011; 7: 26. doi: 10.1186/1746-6148-7-26 21649893

17. Franke J, Abs V, Zizzadoro C, Abraham G. Comparative study of the effects of fetal bovine serum versus horse serum on growth and differentiation of primary equine bronchial fibroblasts. BMC Vet Res. 2014; 10: 119. doi: 10.1186/1746-6148-10-119 24886635

18. Shibeshi W, Abraham G, Kneuer C, Ellenberger C, Seeger J, Schoon H, et al. Isolation and culture of primary equine tracheal epithelial cells. In Vitro Cell Dev Biol Anim. 2008; 44: 179–184. doi: 10.1007/s11626-008-9099-8 18594938

19. Moll R, Divo M, Langbein L. The human keratins: biology and pathology. Histochem Cell Biol. 2008; 129: 705–733. doi: 10.1007/s00418-008-0435-6 18461349

20. Holgate ST, Holloway J, Wilson S, Bucchieri F, Puddicombe S, Davies DE. Epithelial-mesenchymal communication in the pathogenesis of chronic asthma. Proc Am Thorac Soc. 2004; 1: 93–98. doi: 10.1513/pats.2306034 16113419

21. Goto Y, Noguchi Y, Nomura A, Sakamoto T, Ishii Y, Bitoh S, et al. In vitro reconstitution of the tracheal epithelium. Am J Respir Cell Mol Biol. 1999; 20: 312–318. doi: 10.1165/ajrcmb.20.2.3062 9922223

22. Osei ET, Noordhoek JA, Hackett TL, Spanjer AI, Postma DS, Timens W, Brandsma CA, Heijink IH. Interleukin-1α drives the dysfunctional cross-talk of the airway epithelium and lung fibroblasts in COPD. Eur Respir J. 2016; 48: 359–369. doi: 10.1183/13993003.01911-2015 27418555

23. Bogdanowicz DR, Lu HH. Studying cell-cell communication in co-culture. Biotechnol J. 2013; 8: 395–396. doi: 10.1002/biot.201300054 23554248

24. Papazian D, Würtzen PA, Hansen SW. Polarized Airway Epithelial Models for Immunological Co-Culture Studies. Int Arch Allergy Immunol. 2016; 170: 1–21. doi: 10.1159/000445833 27240620

25. Crosby LM, Waters CM. Epithelial repair mechanisms in the lung. Am J Physiol Lung Cell Mol Physiol. 2010; 298: L715–L731. doi: 10.1152/ajplung.00361.2009 20363851

26. Nolte SV, Xu W, Rennekampff HO, Rodemann HP. Diversity of fibroblasts—a review on implications for skin tissue engineering.Cells Tissues Organs 2008; 187: 165–76. doi: 10.1159/000111805 18042973


Článek vyšel v časopise

PLOS One


2019 Číslo 11
Nejčtenější tento týden
Nejčtenější v tomto čísle
Kurzy

Zvyšte si kvalifikaci online z pohodlí domova

KOST
Koncepce osteologické péče pro gynekology a praktické lékaře
nový kurz
Autoři: MUDr. František Šenk

Sekvenční léčba schizofrenie
Autoři: MUDr. Jana Hořínková

Hypertenze a hypercholesterolémie – synergický efekt léčby
Autoři: prof. MUDr. Hana Rosolová, DrSc.

Svět praktické medicíny 5/2023 (znalostní test z časopisu)

Imunopatologie? … a co my s tím???
Autoři: doc. MUDr. Helena Lahoda Brodská, Ph.D.

Všechny kurzy
Kurzy Podcasty Doporučená témata Časopisy
Přihlášení
Zapomenuté heslo

Zadejte e-mailovou adresu, se kterou jste vytvářel(a) účet, budou Vám na ni zaslány informace k nastavení nového hesla.

Přihlášení

Nemáte účet?  Registrujte se

#ADS_BOTTOM_SCRIPTS#