Isolation, culture, and characterization of chicken intestinal epithelial cells

BMC Molecular and Cell Biology

Federico Ghiselli¹  Martina Felici¹  Andrea Piva²  Ester Grilli³  Giovanni Tosi⁴  Maria Parigi⁴  Laura Fiorentini⁴  Paola Massi⁴  Barbara Rossi⁵ 

[1]DIMEVET, University of Bologna, Via Tolara di Sopra, 50, Ozzano dell’Emilia, 40064, Bologna, BO, Italy

[2]DIMEVET, University of Bologna, Via Tolara di Sopra, 50, Ozzano dell’Emilia, 40064, Bologna, BO, Italy

[3]Vetagro S.p.A., Via Ignazio Porro, 2, 42124, Reggio Emilia, RE, Italy

[4]DIMEVET, University of Bologna, Via Tolara di Sopra, 50, Ozzano dell’Emilia, 40064, Bologna, BO, Italy

[5]Vetagro, Inc., 116 W. Jackson Blwd., Suite #320, 60604, Chicago, IL, USA

[6]Istituto Zooprofilattico Sperimentale Della Lombardia e Dell’Emilia Romagna, Sede Territoriale di Forlì, Via Don Eugenio Servadei, 47122, Forlì, FC, Italy

[7]Vetagro S.p.A., Via Ignazio Porro, 2, 42124, Reggio Emilia, RE, Italy

关键词: Chicken;    Enterocyte;    Immunofluorescence;    Embryo;    Primary culture;   

DOI  :  10.1186/s12860-021-00349-7

来源: Springer

【 摘 要 】

BackgroundEnterocytes exert an absorptive and protective function in the intestine, and they encounter many different challenging factors such as feed, bacteria, and parasites. An intestinal epithelial in vitro model can help to understand how enterocytes are affected by these factors and contribute to the development of strategies against pathogens.ResultsThe present study describes a novel method to culture and maintain primary chicken enterocytes and their characterization by immunofluorescence and biomolecular approaches. Starting from 19-day-old chicken embryos it was possible to isolate viable intestinal cell aggregates that can expand and produce a self-maintaining intestinal epithelial cell population that survives until 12 days in culture. These cells resulted positive in immunofluorescence to Cytokeratin 18, Zonula occludens 1, Villin, and Occludin that are common intestinal epithelial markers, and negative to Vimentin that is expressed by endothelial cells. Cells were cultured also on Transwell® permeable supports and trans-epithelial electrical resistance, was measured. This value gradually increased reaching 64 Ω*cm2 7 days after seeding and it remained stable until day 12.ConclusionsBased on these results it was confirmed that it is possible to isolate and maintain chicken intestinal epithelial cells in culture and that they can be suitable as in vitro intestinal model for further studies.

【 授权许可】

CC BY