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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Published in | International journal of molecular sciences Vol. 17; no. 10; p. 1655 |
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Abstract | 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
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. |
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AbstractList | 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. 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 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. 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. |
Author | Hammer, Susanne C Mohr, Annika Brenig, Bertram Ngezahayo, Anaclet Hennecke, Silvia Nolte, Ingo Becker, Annegret Lüder Ripoli, Florenza Rateitschak, Katja Murua Escobar, Hugo Junginger, Johannes Hewicker-Trautwein, Marion |
AuthorAffiliation | 3 Institute of Biophysics, Leibniz University Hannover, Herrenhäuser Straße 2, 30419 Hannover, Germany; a.becker@biophysik.uni-hannover.de (A.B.); ngezahayo@biophysik.uni-hannover.de (A.N.) 4 Institute for Bioinformatics, University Medicine Greifswald, Walther-Rathenau-Str. 48, 17475 Greifswald, Germany; katja.rateitschak@uni-greifswald.de 5 Institute of Veterinary Medicine, Georg-August-University Göttingen, Burckhardtweg 2, 37077 Göttingen, Germany; shennec@gwdg.de (S.H.); bbrenig@gwdg.de (B.B.) 1 Small Animal Clinic, University of Veterinary Medicine Hannover, Bünteweg 9, 30559 Hannover, Germany; schammer@tiho-hannover.de (S.C.H.); annika.mohr@tiho-hannover.de (A.M.); florenza@ripoli.com.br (F.L.R.); hugo.murua.escobar@med.uni-rostock.de (H.M.E.) 2 Division of Medicine, Haematology, Oncology and Palliative Medicine, University of Rostock, Ernst-Heydemann-Str. 6, 18055 Rostock, Germany 7 Center for Systems Neuroscience (ZSN) Hannover, University of Veterinary Medicine Hannover, Bünteweg |
AuthorAffiliation_xml | – name: 6 Department of Pathology, University of Veterinary Medicine Hannover, Bünteweg 17, 30559 Hannover, Germany; johannes.junginger@tiho-hannover.de (J.J.); marion.hewicker-trautwein@tiho-hannover.de (M.H.-T.) – name: 4 Institute for Bioinformatics, University Medicine Greifswald, Walther-Rathenau-Str. 48, 17475 Greifswald, Germany; katja.rateitschak@uni-greifswald.de – name: 7 Center for Systems Neuroscience (ZSN) Hannover, University of Veterinary Medicine Hannover, Bünteweg 17, 30559 Hannover, Germany – name: 2 Division of Medicine, Haematology, Oncology and Palliative Medicine, University of Rostock, Ernst-Heydemann-Str. 6, 18055 Rostock, Germany – name: 1 Small Animal Clinic, University of Veterinary Medicine Hannover, Bünteweg 9, 30559 Hannover, Germany; schammer@tiho-hannover.de (S.C.H.); annika.mohr@tiho-hannover.de (A.M.); florenza@ripoli.com.br (F.L.R.); hugo.murua.escobar@med.uni-rostock.de (H.M.E.) – name: 3 Institute of Biophysics, Leibniz University Hannover, Herrenhäuser Straße 2, 30419 Hannover, Germany; a.becker@biophysik.uni-hannover.de (A.B.); ngezahayo@biophysik.uni-hannover.de (A.N.) – name: 5 Institute of Veterinary Medicine, Georg-August-University Göttingen, Burckhardtweg 2, 37077 Göttingen, Germany; shennec@gwdg.de (S.H.); bbrenig@gwdg.de (B.B.) |
Author_xml | – sequence: 1 givenname: Susanne C surname: Hammer fullname: Hammer, Susanne C email: schammer@tiho-hannover.de, schammer@tiho-hannover.de organization: Division of Medicine, Haematology, Oncology and Palliative Medicine, University of Rostock, Ernst-Heydemann-Str. 6, 18055 Rostock, Germany. schammer@tiho-hannover.de – sequence: 2 givenname: Annegret surname: Becker fullname: Becker, Annegret email: a.becker@biophysik.uni-hannover.de organization: Institute of Biophysics, Leibniz University Hannover, Herrenhäuser Straße 2, 30419 Hannover, Germany. a.becker@biophysik.uni-hannover.de – sequence: 3 givenname: Katja surname: Rateitschak fullname: Rateitschak, Katja email: katja.rateitschak@uni-greifswald.de organization: Institute for Bioinformatics, University Medicine Greifswald, Walther-Rathenau-Str. 48, 17475 Greifswald, Germany. katja.rateitschak@uni-greifswald.de – sequence: 4 givenname: Annika surname: Mohr fullname: Mohr, Annika email: annika.mohr@tiho-hannover.de, annika.mohr@tiho-hannover.de organization: Division of Medicine, Haematology, Oncology and Palliative Medicine, University of Rostock, Ernst-Heydemann-Str. 6, 18055 Rostock, Germany. annika.mohr@tiho-hannover.de – sequence: 5 givenname: Florenza surname: Lüder Ripoli fullname: Lüder Ripoli, Florenza email: florenza@ripoli.com.br, florenza@ripoli.com.br organization: Division of Medicine, Haematology, Oncology and Palliative Medicine, University of Rostock, Ernst-Heydemann-Str. 6, 18055 Rostock, Germany. florenza@ripoli.com.br – sequence: 6 givenname: Silvia surname: Hennecke fullname: Hennecke, Silvia email: shennec@gwdg.de organization: Institute of Veterinary Medicine, Georg-August-University Göttingen, Burckhardtweg 2, 37077 Göttingen, Germany. shennec@gwdg.de – sequence: 7 givenname: Johannes surname: Junginger fullname: Junginger, Johannes email: johannes.junginger@tiho-hannover.de organization: Department of Pathology, University of Veterinary Medicine Hannover, Bünteweg 17, 30559 Hannover, Germany. johannes.junginger@tiho-hannover.de – sequence: 8 givenname: Marion surname: Hewicker-Trautwein fullname: Hewicker-Trautwein, Marion email: marion.hewicker-trautwein@tiho-hannover.de organization: Department of Pathology, University of Veterinary Medicine Hannover, Bünteweg 17, 30559 Hannover, Germany. marion.hewicker-trautwein@tiho-hannover.de – sequence: 9 givenname: Bertram surname: Brenig fullname: Brenig, Bertram email: bbrenig@gwdg.de organization: Institute of Veterinary Medicine, Georg-August-University Göttingen, Burckhardtweg 2, 37077 Göttingen, Germany. bbrenig@gwdg.de – sequence: 10 givenname: Anaclet surname: Ngezahayo fullname: Ngezahayo, Anaclet email: ngezahayo@biophysik.uni-hannover.de, ngezahayo@biophysik.uni-hannover.de organization: Center for Systems Neuroscience (ZSN) Hannover, University of Veterinary Medicine Hannover, Bünteweg 17, 30559 Hannover, Germany. ngezahayo@biophysik.uni-hannover.de – sequence: 11 givenname: Ingo surname: Nolte fullname: Nolte, Ingo email: ingo.nolte@tiho-hannover.de organization: Small Animal Clinic, University of Veterinary Medicine Hannover, Bünteweg 9, 30559 Hannover, Germany. ingo.nolte@tiho-hannover.de – sequence: 12 givenname: Hugo surname: Murua Escobar fullname: Murua Escobar, Hugo email: hugo.murua.escobar@med.uni-rostock.de, hugo.murua.escobar@med.uni-rostock.de organization: Division of Medicine, Haematology, Oncology and Palliative Medicine, University of Rostock, Ernst-Heydemann-Str. 6, 18055 Rostock, Germany. hugo.murua.escobar@med.uni-rostock.de |
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Keywords | canine cell culture cell lines mammary neoplasias claudin marker expression |
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Title | Longitudinal Claudin Gene Expression Analyses in Canine Mammary Tissues and Thereof Derived Primary Cultures and Cell Lines |
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