Protein profiles in the transfected oviductal secreting cells of laying hen (Gallus gallus domesticus)
ABSTRACT: Due to the intensive development of novel biopharming applications, there is a need for the in vitro verification models prior to in vivo testing. Laying hen has been already applied as an animal bioreactor to produce the therapeutical enzyme in a rare disease called lysosomal acid lipase...
| Main Authors: | , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2024-03-01
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| Series: | Poultry Science |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S0032579123008246 |
| _version_ | 1827338989910097920 |
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| author | Mengjun Wu Giuseppe Maiorano Katarzyna Stadnicka |
| author_facet | Mengjun Wu Giuseppe Maiorano Katarzyna Stadnicka |
| author_sort | Mengjun Wu |
| collection | DOAJ |
| description | ABSTRACT: Due to the intensive development of novel biopharming applications, there is a need for the in vitro verification models prior to in vivo testing. Laying hen has been already applied as an animal bioreactor to produce the therapeutical enzyme in a rare disease called lysosomal acid lipase deficiency. In this study, we aimed to verify how the proteome of the transfected oviduct epithelial cells would be affected by genetic nonviral modification with the human exogene. The study was based on a previously developed method to cultivate chicken oviduct epithelial cells (COEC). The typical characteristics of the COEC epithelial cells were retained across the experiments. The mean efficiency of nucleofection ranged from 2.6 to 19.7% depending on the cells’ isolation and location in the oviduct (upper, infundibulum site, or magnum). The PCR confirmed the incorporation of human interferon alpha2a (hIFNα2a) exogene into the nucleofected COEC but, the production of hIFNα2a protein did not exceed the detection level in this study. The ovalbumin protein was detected in the nontransfected and transfected COEC, which confirmed the normal secreting functions of the cells subject to modification. Proteomic analysis revealed an increase in abundance of the cell adhesion molecules and collagen molecules after introducing gene under ovalbumin promoter. According to the bioinformatic analyses there was a limited negative impact of transfection on cells, and the normal biochemical pathways were not severely disordered. In conclusion, the observations provide new knowledge about the proteomic profile of the manipulated COEC with regard to the retained normal functionality of the cells, which can be informative for avian biopharma research. |
| first_indexed | 2024-03-07T19:43:36Z |
| format | Article |
| id | doaj.art-8d82a6d09cdd4a6997bdba87cd679a73 |
| institution | Directory Open Access Journal |
| issn | 0032-5791 |
| language | English |
| last_indexed | 2024-03-07T19:43:36Z |
| publishDate | 2024-03-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Poultry Science |
| spelling | doaj.art-8d82a6d09cdd4a6997bdba87cd679a732024-02-29T05:17:07ZengElsevierPoultry Science0032-57912024-03-011033103305Protein profiles in the transfected oviductal secreting cells of laying hen (Gallus gallus domesticus)Mengjun Wu0Giuseppe Maiorano1Katarzyna Stadnicka2Department of Agricultural, Environmental and Food Sciences, University of Molise, 86100 Campobasso, ItalyDepartment of Agricultural, Environmental and Food Sciences, University of Molise, 86100 Campobasso, ItalyFaculty of Health Sciences, Ludwik Rydygier Collegium Medicum in Bydgoszcz, Nicolaus Copernicus University in Toruń, 85-821 Bydgoszcz, Poland; Corresponding author:ABSTRACT: Due to the intensive development of novel biopharming applications, there is a need for the in vitro verification models prior to in vivo testing. Laying hen has been already applied as an animal bioreactor to produce the therapeutical enzyme in a rare disease called lysosomal acid lipase deficiency. In this study, we aimed to verify how the proteome of the transfected oviduct epithelial cells would be affected by genetic nonviral modification with the human exogene. The study was based on a previously developed method to cultivate chicken oviduct epithelial cells (COEC). The typical characteristics of the COEC epithelial cells were retained across the experiments. The mean efficiency of nucleofection ranged from 2.6 to 19.7% depending on the cells’ isolation and location in the oviduct (upper, infundibulum site, or magnum). The PCR confirmed the incorporation of human interferon alpha2a (hIFNα2a) exogene into the nucleofected COEC but, the production of hIFNα2a protein did not exceed the detection level in this study. The ovalbumin protein was detected in the nontransfected and transfected COEC, which confirmed the normal secreting functions of the cells subject to modification. Proteomic analysis revealed an increase in abundance of the cell adhesion molecules and collagen molecules after introducing gene under ovalbumin promoter. According to the bioinformatic analyses there was a limited negative impact of transfection on cells, and the normal biochemical pathways were not severely disordered. In conclusion, the observations provide new knowledge about the proteomic profile of the manipulated COEC with regard to the retained normal functionality of the cells, which can be informative for avian biopharma research.http://www.sciencedirect.com/science/article/pii/S0032579123008246henchicken oviduct epithelial cellsnonviral transfectionproteomicssecretion |
| spellingShingle | Mengjun Wu Giuseppe Maiorano Katarzyna Stadnicka Protein profiles in the transfected oviductal secreting cells of laying hen (Gallus gallus domesticus) Poultry Science hen chicken oviduct epithelial cells nonviral transfection proteomics secretion |
| title | Protein profiles in the transfected oviductal secreting cells of laying hen (Gallus gallus domesticus) |
| title_full | Protein profiles in the transfected oviductal secreting cells of laying hen (Gallus gallus domesticus) |
| title_fullStr | Protein profiles in the transfected oviductal secreting cells of laying hen (Gallus gallus domesticus) |
| title_full_unstemmed | Protein profiles in the transfected oviductal secreting cells of laying hen (Gallus gallus domesticus) |
| title_short | Protein profiles in the transfected oviductal secreting cells of laying hen (Gallus gallus domesticus) |
| title_sort | protein profiles in the transfected oviductal secreting cells of laying hen gallus gallus domesticus |
| topic | hen chicken oviduct epithelial cells nonviral transfection proteomics secretion |
| url | http://www.sciencedirect.com/science/article/pii/S0032579123008246 |
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