Longitudinal Claudin Gene Expression Analyses in Canine Mammary Tissues and Thereof Derived Primary Cultures and Cell Lines

Human and canine mammary tumours show partial claudin expression deregulations. Further, claudins have been used for directed therapeutic approaches. However, the development of claudin targeting approaches requires stable claudin expressing cell lines. This study reports the establishment and chara...

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Main Authors:	Susanne C. Hammer, Annegret Becker, Katja Rateitschak, Annika Mohr, Florenza Lüder Ripoli, Silvia Hennecke, Johannes Junginger, Marion Hewicker-Trautwein, Bertram Brenig, Anaclet Ngezahayo, Ingo Nolte, Hugo Murua Escobar
Format:	Article
Language:	English
Published:	MDPI AG 2016-09-01
Series:	International Journal of Molecular Sciences
Subjects:	claudin mammary neoplasias canine cell lines cell culture marker expression
Online Access:	http://www.mdpi.com/1422-0067/17/10/1655

id	doaj-24cf8bae38e142ac992159d5585f8cbd
record_format	Article
spelling	doaj-24cf8bae38e142ac992159d5585f8cbd2020-11-24T23:53:23ZengMDPI AGInternational Journal of Molecular Sciences1422-00672016-09-011710165510.3390/ijms17101655ijms17101655Longitudinal Claudin Gene Expression Analyses in Canine Mammary Tissues and Thereof Derived Primary Cultures and Cell LinesSusanne C. Hammer0Annegret Becker1Katja Rateitschak2Annika Mohr3Florenza Lüder Ripoli4Silvia Hennecke5Johannes Junginger6Marion Hewicker-Trautwein7Bertram Brenig8Anaclet Ngezahayo9Ingo Nolte10Hugo Murua Escobar11Small Animal Clinic, University of Veterinary Medicine Hannover, Bünteweg 9, 30559 Hannover, GermanyInstitute of Biophysics, Leibniz University Hannover, Herrenhäuser Straße 2, 30419 Hannover, GermanyInstitute for Bioinformatics, University Medicine Greifswald, Walther-Rathenau-Str. 48, 17475 Greifswald, GermanySmall Animal Clinic, University of Veterinary Medicine Hannover, Bünteweg 9, 30559 Hannover, GermanySmall Animal Clinic, University of Veterinary Medicine Hannover, Bünteweg 9, 30559 Hannover, GermanyInstitute of Veterinary Medicine, Georg-August-University Göttingen, Burckhardtweg 2, 37077 Göttingen, GermanyDepartment of Pathology, University of Veterinary Medicine Hannover, Bünteweg 17, 30559 Hannover, GermanyDepartment of Pathology, University of Veterinary Medicine Hannover, Bünteweg 17, 30559 Hannover, GermanyInstitute of Veterinary Medicine, Georg-August-University Göttingen, Burckhardtweg 2, 37077 Göttingen, GermanyInstitute of Biophysics, Leibniz University Hannover, Herrenhäuser Straße 2, 30419 Hannover, GermanySmall Animal Clinic, University of Veterinary Medicine Hannover, Bünteweg 9, 30559 Hannover, GermanySmall Animal Clinic, University of Veterinary Medicine Hannover, Bünteweg 9, 30559 Hannover, GermanyHuman and canine mammary tumours show partial claudin expression deregulations. Further, claudins have been used for directed therapeutic approaches. However, the development of claudin targeting approaches requires stable claudin expressing cell lines. This study reports the establishment and characterisation of canine mammary tissue derived cell lines, analysing longitudinally the claudin-1, -3, -4 and -7 expressions in original tissue samples, primary cultures and developed cell lines. Primary cultures were derived from 17 canine mammary tissues: healthy, lobular hyperplasia, simple adenoma, complex adenoma, simple tubular carcinoma, complex carcinoma, carcinoma arising in a benign mixed tumour and benign mixed tissue. Cultivation was performed, if possible, until passage 30. Claudin mRNA and protein expressions were analysed by PCR, QuantiGene Plex Assay, immunocytochemistry and immunofluorescence. Further, cytokeratin expression was analysed immunocytochemically. Cultivation resulted in 11 established cell lines, eight showing epithelial character. In five of the early passages the claudin expressions decreased compared to the original tissues. In general, claudin expressions were diminished during cultivation. Three cell lines kept longitudinally claudin, as well as epithelial marker expressions, representing valuable tools for the development of claudin targeted anti-tumour therapies.http://www.mdpi.com/1422-0067/17/10/1655claudinmammary neoplasiascaninecell linescell culturemarker expression
collection	DOAJ
language	English
format	Article
sources	DOAJ
author	Susanne C. Hammer Annegret Becker Katja Rateitschak Annika Mohr Florenza Lüder Ripoli Silvia Hennecke Johannes Junginger Marion Hewicker-Trautwein Bertram Brenig Anaclet Ngezahayo Ingo Nolte Hugo Murua Escobar
spellingShingle	Susanne C. Hammer Annegret Becker Katja Rateitschak Annika Mohr Florenza Lüder Ripoli Silvia Hennecke Johannes Junginger Marion Hewicker-Trautwein Bertram Brenig Anaclet Ngezahayo Ingo Nolte Hugo Murua Escobar Longitudinal Claudin Gene Expression Analyses in Canine Mammary Tissues and Thereof Derived Primary Cultures and Cell Lines International Journal of Molecular Sciences claudin mammary neoplasias canine cell lines cell culture marker expression
author_facet	Susanne C. Hammer Annegret Becker Katja Rateitschak Annika Mohr Florenza Lüder Ripoli Silvia Hennecke Johannes Junginger Marion Hewicker-Trautwein Bertram Brenig Anaclet Ngezahayo Ingo Nolte Hugo Murua Escobar
author_sort	Susanne C. Hammer
title	Longitudinal Claudin Gene Expression Analyses in Canine Mammary Tissues and Thereof Derived Primary Cultures and Cell Lines
title_short	Longitudinal Claudin Gene Expression Analyses in Canine Mammary Tissues and Thereof Derived Primary Cultures and Cell Lines
title_full	Longitudinal Claudin Gene Expression Analyses in Canine Mammary Tissues and Thereof Derived Primary Cultures and Cell Lines
title_fullStr	Longitudinal Claudin Gene Expression Analyses in Canine Mammary Tissues and Thereof Derived Primary Cultures and Cell Lines
title_full_unstemmed	Longitudinal Claudin Gene Expression Analyses in Canine Mammary Tissues and Thereof Derived Primary Cultures and Cell Lines
title_sort	longitudinal claudin gene expression analyses in canine mammary tissues and thereof derived primary cultures and cell lines
publisher	MDPI AG
series	International Journal of Molecular Sciences
issn	1422-0067
publishDate	2016-09-01
description	Human and canine mammary tumours show partial claudin expression deregulations. Further, claudins have been used for directed therapeutic approaches. However, the development of claudin targeting approaches requires stable claudin expressing cell lines. This study reports the establishment and characterisation of canine mammary tissue derived cell lines, analysing longitudinally the claudin-1, -3, -4 and -7 expressions in original tissue samples, primary cultures and developed cell lines. Primary cultures were derived from 17 canine mammary tissues: healthy, lobular hyperplasia, simple adenoma, complex adenoma, simple tubular carcinoma, complex carcinoma, carcinoma arising in a benign mixed tumour and benign mixed tissue. Cultivation was performed, if possible, until passage 30. Claudin mRNA and protein expressions were analysed by PCR, QuantiGene Plex Assay, immunocytochemistry and immunofluorescence. Further, cytokeratin expression was analysed immunocytochemically. Cultivation resulted in 11 established cell lines, eight showing epithelial character. In five of the early passages the claudin expressions decreased compared to the original tissues. In general, claudin expressions were diminished during cultivation. Three cell lines kept longitudinally claudin, as well as epithelial marker expressions, representing valuable tools for the development of claudin targeted anti-tumour therapies.
topic	claudin mammary neoplasias canine cell lines cell culture marker expression
url	http://www.mdpi.com/1422-0067/17/10/1655
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Longitudinal Claudin Gene Expression Analyses in Canine Mammary Tissues and Thereof Derived Primary Cultures and Cell Lines

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