Establishment and characterization of a cell line from canine mammary gland tumor

• Main Authors: Hou-You Su, 蘇厚有
• Other Authors: Min-Liang Wong
• Format: Others
• Language: zh-TW
• Published: 2001
• Online Access: http://ndltd.ncl.edu.tw/handle/82066803775421543928

碩士 === 國立中興大學 === 獸醫學系 === 89 === Mammary gland tumors are the most common types of neoplasms in bitch. According to histopathologic classification, the percentages of malignant tumors are about 50 %, and mixed type mammary gland tumors are usually developed. Because of the mammary gland is consisted of multiple cell types and forms a complex histogenesis and pathologic change, so that it is a difficult to research in vitro and vivo. If a cell line can be established from canine mammary gland tumor, it may be useful for the research of cell proliferation as well as for the development of anti-cancer reagent. Initially we obtained the mammary gland tumor from the 6 years old patient in the Veterinary Hospital of Chung-Hsing University, and isolated the cell from tumor. Histopathologic diagnosis of the tumor revealed a malignant mixed mammary gland tumor. We have successfully established a cell line, which was named DMGT. The DMGT cells were cultured in flask showing elongated spindle-shape and polygonal cell type. When they were grown to high density, the cell became multiple cluster, stellate-form, and arranged to cross-link. We further analyzed the properties of the cell line by immunocytochemistry. The immunocytochemistry staining results indicated the cells are keratin postive, mucin 1 postive, and vimentin negtive. So we concluded the cell line was a epithelial origin. In the MTT proliferation rate test, the doubling time of DMGT cells was about 48 h. Besides, to investigate the transforming characteristic of DMGT cells, we cultured them in soft agar. After two weeks, the cell focus was formed, and the MTT test demonstrated the cell colonies were viability. In malignancy test, we injected subcutaneously the DMGT cells into the abdomen of SCID mice and into the back of nude mice. About a month later, we observed the mass formation on inocultated site both in SCID mice and in nude mice. The mouse tumor masses were excised and histopathologic examination was performed. Islands of poor differentiated spindle-shaped cells were observed in SCID mouse tissue. Taken together, we established a transformed cell line (DMGT) from a canine mammary tumor, and the immunohistochemical property and the tumorigenecity of DMGT cells were studied.