Newborn pig trachea cell line cultured in air-liquid interface conditions allows a partial in vitro representation of the porcine upper airway tissue

BMC Cell Biology

François Meurens¹  Gaëlle Simon³  Pierre-Yves Sizaret⁴  Michel Olivier²  Mario Delgado-Ortega² 

[1]Vaccine and Infectious Disease Organization-InterVac, University of Saskatchewan, 120 Veterinary Road, Saskatoon S7N 5E3 Saskatchewan, Canada

[2]UMR1282 Infectiologie et Santé Publique, Université François Rabelais, Tours 37000, France

[3]European University of Brittany, Rennes 35000, France

[4]Département des microscopies, plate-forme R.I.O de microscopie électronique, Université François Rabelais, Tours 37000, France

关键词: Trachea;    Air-liquid interface;    Differentiation;    Epithelial cell;    Pig;   

Others  :  854980
DOI  :  10.1186/1471-2121-15-14

 received in 2013-12-17, accepted in 2014-04-23,  发布年份 2014

【 摘 要 】

Background

The domestic pig is an excellent animal model to study human microbial diseases due to its similarity to humans in terms of anatomy, physiology, and genetics. We assessed the suitability of an in vitro air-liquid interface (ALI) culture system for newborn pig trachea (NPTr) cells as a practical tool for analyzing the immune response of respiratory epithelial cells to aggressors. This cell line offers a wide microbial susceptibility spectrum to both viruses and bacteria. The purpose of our study was to evaluate and characterize diverse aspects of cell differentiation using different culture media. After the NPTr cells reached confluence, the apical medium was removed and the cells were fed by medium from the basal side.

Results

We assessed the cellular layer’s capacity to polarize and differentiate in ALI conditions. Using immunofluorescence and electronic microscopy we evaluated the presence of goblet and ciliated cells, the epithelial junction organization, and the transepithelial electrical resistance. We found that the cellular layer develops a variable density of mucus producing cells and acquires a transepithelial resistance. We also identified increased development of cellular junctions over the culture period. Finally, we observed variable expression of transcripts associated to proteins such as keratin 8, mucins (MUC1, MUC2, and MUC4), occludin, and villin 1.

Conclusions

The culture of NPTr cells in ALI conditions allows a partial in vitro representation of porcine upper airway tissue that could be used to investigate some aspects of host/respiratory pathogen interactions.

【 授权许可】

   
2014 Delgado-Ortega et al.; licensee BioMed Central Ltd.

【 图 表 】

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