Establishment and Characterization of Canine Mammary Gland Cancer Cell Lines

개 자연발생 유선종양 2종의 세포주 확립 및 특성분석

  • Lee, Sun-Tae (Department of Veterinary Surgery, College of Veterinary Medicine, Seoul National University) ;
  • Kweon, Oh-Kyeong (Department of Veterinary Surgery, College of Veterinary Medicine, Seoul National University) ;
  • Kim, Wan-Hee (Department of Veterinary Surgery, College of Veterinary Medicine, Seoul National University)
  • Accepted : 2010.03.31
  • Published : 2010.06.30

Abstract

Two cell lines derived from spontaneous canine mammary gland tumors were established and characterized. Mammary gland tumors from 9 years old pug and 9 years old toy-poodle dogs were collected by aseptic surgical resection and primary culture was performed. The histopathologic examination of tumors revealed adenocarcinoma and complex carcinoma and two dogs died from metastasis of the tumors. The tumor cells were subcultured over 60 times for more than 1 year and morphological consistency maintained. Light microscopic examination, growth curve, doubling time calculation, xenotransplantation to female nude mice, immunohistochemistry for wide spectrum keratin, vimentin, $\alpha$-smooth muscle actin and cytokeratin 8 was performed for characterization. The cell lines exhibited polygonal, elongated cell shape and cytoplasmic bridge and doubling time of 47.1 hrs and 18.6 hrs, respectively. Subcutaneous xenotransplantation to nude mice of the cells produced localized palpable mass within 4 weeks in 4 of 5 and 5 of 5 nude mice, respectively. In immunohistochemical examination one cell line showed strong positive against wide spectrum keratin and cytokeratin 8 and the other cell line showed strong positive against smooth muscle actin and cytokeratin 8. Additional characterization would be possible by investigator's needs and the cell lines may be useful for in vivo and in vitro studies of canine mammary tumor and adjuvant therapies.

개에서 자연발생한 유선종양으로부터 채취, 배양된 세포주 2개를 확립하고 특성을 분석하였다. 9세 령의 퍼그 종 개와 동일연령의 토이푸들 종의 개에 발생한 종양을 무균 상태를 유지하여 수술적으로 채취한 후 primary culture를 실시하였다. 두 종양의 조직병리검사에서는 각각 선암종과 혼합암종이 진단되었으며, 이후 두 마리 모두 전이로 인해 폐사하였다. 배양된 종양세포는 1년 이상의 기간 동안 60회 이상 계대를 반복하면서 형태학적으로 일관성을 유지하였고, 특성분석을 위해 광학현미경검사, 성장곡선 산출, 배가 시간 계산, 누드 마우스에 이종이식, 면역조직 화학검사를 실시하였다. 각 세포주는 다각형의 긴 세포형태를 보였으며, 세포질 연결을 형성하였으며, 배가 시간은 각각 47.1 시간과 18.6 시간이었다. 암컷 누드마우스의 등 부위에 피하이식 후 4주 이내에 10마리 중 9마리에서 촉진이 가능한 종괴의 형성이 확인되었으며, 면역조직화학검사 시 한 세포주에서는 keratin과 cytokeratin 8에서 다른 세포주에서는 smooth muscle actin과 cytokeratin 8에서 강한 염색성이 확인되었다. 두 세포주는 개의 유선종양의 시험관내, 체내 연구에 있어 모두 유용하게 사용될 수 있을 것으로 생각된다.

Keywords

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