Abstract
A novel cell line of canine medullary thyroid carcinoma (MTC) was established from the neck mass, diagnosed histopathologically and immunohistochemically as ectopic MTC. The neoplastic cells arranging trabecular structures were characterized as pleomorphic cells with eosinophilic cytoplasm and nucleus, containing often clear nucleolus. These tumor cells were immuno-positive for calcitonin gene–related protein (CGRP), somatostatin, and chromogranin A. In addition, 8th passaged cultured cells were also immuno-positive for CGRP, somatostatin, and chromogranin A. The cloned tumor cells showed logarithmic cell growth with a doubling time of 33.3 h. From the results of DNA sequencing of rearranged during transfection (RET) proto-oncogene, the cloned tumor cells had four single base substitution, including exon 5 codon 82, exon 16 codon 750, exon 17 codon 777, and exon 24 codon 1085, all of which were single nucleotide polymorphism reported in RET gene of dogs. After the xenotransplantation into severe combined immunodeficiency (SCID) mice, the cloned cells showed tumorigenicity potentials. The morphological and immunohistochemical features of the xenotransplanted tumor were almost in conformity with those of the original tumor, including positive immunoreactivity for calcitonin, CGRP, and chromogranin A. To our knowledge, this is the first report of canine MTC cell line, which provides useful in vitro tool for understanding oncogenic mechanism and pathophysiological state of MTC in dogs.
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All procedures performed in studies involving animals were in accordance with the ethical standards of the institution or practice at which the studies were conducted.
I already described in manuscript, line 141–150, “The injected mice were housed in a cage with free access to water and a standard purified rodent growth diet (AIN-93G, Oriental Yeast, Tokyo, Japan). When the mass was palpable at the injection site, the tumor volume (length × width × height) was measured every day. At the 57th day after the injection, the mice were euthanized with isoflurane and the tumor was resected for histopathological and immunohistochemical examination. All animal experiments were approved by the Committee of Animal Experiments, Graduate School of Agricultural and Life Sciences, the University of Tokyo (approve number P17-017).
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Okano, K., Uematsu, Y., Nibe, K. et al. Establishment and characterization of a novel cell line of medullary thyroid carcinoma from a dog. In Vitro Cell.Dev.Biol.-Animal 55, 559–566 (2019). https://doi.org/10.1007/s11626-019-00339-2
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DOI: https://doi.org/10.1007/s11626-019-00339-2