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

authored by

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

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 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.

Organisation(s)

Institute of Cell Biology and Biophysics

External Organisation(s)

University of Veterinary Medicine of Hannover, Foundation
University of Greifswald
University of Göttingen
Center for Systems Neuroscience Hannover (ZSN)

Type

Article

Journal

International Journal of Molecular Sciences

Volume

17

ISSN

1422-0067

Publication date

29.09.2016

Publication status

Published

Peer reviewed

Yes

ASJC Scopus subject areas

Molecular Biology, Spectroscopy, Catalysis, Inorganic Chemistry, Computer Science Applications, Physical and Theoretical Chemistry, Organic Chemistry

Sustainable Development Goals

SDG 3 - Good Health and Well-being

Electronic version(s)

https://doi.org/10.3390/ijms17101655 (Access: Open)