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Article

Development of a Pig Mammary Epithelial Cell Culture Model as a Non-Clinical Tool for Studying Epithelial Barrier—A Contribution from the IMI-ConcePTION Project

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Department of Veterinary Medical Sciences, University of Bologna, Ozzano dell’Emilia, 40064 Bologna, Italy
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Health Sciences and Technologies-Interdepartmental Center for Industrial Research (CIRI-SDV), Alma Mater Studiorum—University of Bologna, 40126 Bologna, Italy
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KU Leuven Drug Delivery and Disposition Lab, Department of Pharmaceutical and Pharmacological Sciences, O&N II Herestraat, 49 3000 Leuven, Belgium
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Novartis Pharma AG, Novartis Institutes for BioMedical Research, Werk Klybeck Postfach, CH-4002 Basel, Switzerland
*
Author to whom correspondence should be addressed.
Academic Editors: Katarzyna Stadnicka, Luiz Cordeiro and Adelina Gama
Animals 2021, 11(7), 2012; https://doi.org/10.3390/ani11072012
Received: 13 April 2021 / Revised: 2 June 2021 / Accepted: 2 July 2021 / Published: 5 July 2021
The information about the risks related to the use of medication during breastfeeding is lacking for most commonly used drugs. The ConcePTION project aims to fill this gap using multiple approaches. Within the project, the pig has been selected as the most appropriate in vivo animal model. In agreement with the application of the “3Rs” principle (Replacement, Reduction and Refinement) and international legislations, the present paper reports the establishment of cellular lines of porcine mammary epithelial cells as a valid tool to study the mammary epithelial barrier function in vitro.
The ConcePTION project aims at generating further knowledge about the risks related to the use of medication during breastfeeding, as this information is lacking for most commonly used drugs. Taking into consideration multiple aspects, the pig model has been considered by the consortium as the most appropriate choice. The present research was planned to develop an efficient method for the isolation and culture of porcine Mammary Epithelial Cells (pMECs) to study the mammary epithelial barrier in vitro. Mammary gland tissues were collected at a local slaughterhouse, dissociated and the selected cellular population was cultured, expanded and characterized by morphology, cell cycle analysis and immunophenotyping. Their ability to create a barrier was tested by TEER measurement and sodium fluorescein transport activity. Expression of 84 genes related to drug transporters was evaluated by a PCR array. Our results show that primary cells express epithelial cell markers: CKs, CK18, E-Cad and tight junctions molecules ZO-1 and OCL. All the three pMEC cellular lines were able to create a tight barrier, although with different strengths and kinetics, and express the main ABC and SLC drug transporters. In conclusion, in the present paper we have reported an efficient method to obtain primary pMEC lines to study epithelial barrier function in the pig model. View Full-Text
Keywords: lactation; breastfeeding; mammary gland; pig model; mammary epithelial cells; barrier function lactation; breastfeeding; mammary gland; pig model; mammary epithelial cells; barrier function
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MDPI and ACS Style

Bernardini, C.; La Mantia, D.; Salaroli, R.; Zannoni, A.; Nauwelaerts, N.; Deferm, N.; Ventrella, D.; Bacci, M.L.; Sarli, G.; Bouisset-Leonard, M.; Annaert, P.; Forni, M. Development of a Pig Mammary Epithelial Cell Culture Model as a Non-Clinical Tool for Studying Epithelial Barrier—A Contribution from the IMI-ConcePTION Project. Animals 2021, 11, 2012. https://doi.org/10.3390/ani11072012

AMA Style

Bernardini C, La Mantia D, Salaroli R, Zannoni A, Nauwelaerts N, Deferm N, Ventrella D, Bacci ML, Sarli G, Bouisset-Leonard M, Annaert P, Forni M. Development of a Pig Mammary Epithelial Cell Culture Model as a Non-Clinical Tool for Studying Epithelial Barrier—A Contribution from the IMI-ConcePTION Project. Animals. 2021; 11(7):2012. https://doi.org/10.3390/ani11072012

Chicago/Turabian Style

Bernardini, Chiara, Debora La Mantia, Roberta Salaroli, Augusta Zannoni, Nina Nauwelaerts, Neel Deferm, Domenico Ventrella, Maria Laura Bacci, Giuseppe Sarli, Michele Bouisset-Leonard, Pieter Annaert, and Monica Forni. 2021. "Development of a Pig Mammary Epithelial Cell Culture Model as a Non-Clinical Tool for Studying Epithelial Barrier—A Contribution from the IMI-ConcePTION Project" Animals 11, no. 7: 2012. https://doi.org/10.3390/ani11072012

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