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 inInternational journal of molecular sciences Vol. 17; no. 10; p. 1655
Main Authors Hammer, Susanne C, Becker, Annegret, Rateitschak, Katja, Mohr, Annika, Lüder Ripoli, Florenza, Hennecke, Silvia, Junginger, Johannes, Hewicker-Trautwein, Marion, Brenig, Bertram, Ngezahayo, Anaclet, Nolte, Ingo, Murua Escobar, Hugo
Format Journal Article
LanguageEnglish
Published Switzerland MDPI AG 01.10.2016
MDPI
Subjects
Breasts
canine
cell culture
cell lines
claudin
Gene expression
mammary neoplasias
marker expression
Proteins
Tumors
canine
cell culture
cell lines
mammary neoplasias
claudin
marker expression
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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.
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
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– 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.)
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  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
BackLink https://www.ncbi.nlm.nih.gov/pubmed/27690019$$D View this record in MEDLINE/PubMed
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Keywords canine
cell culture
cell lines
mammary neoplasias
claudin
marker expression
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Snippet Human and canine mammary tumours show partial claudin expression deregulations. Further, claudins have been used for directed therapeutic approaches. However,...
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SubjectTerms Breasts
canine
cell culture
cell lines
claudin
Gene expression
mammary neoplasias
marker expression
Proteins
Tumors
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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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