| Project/Area Number |
61480387
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
Functional basic dentistry
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| Research Institution | Osaka University |
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Principal Investigator |
TAKIGAWA Masaharu Department of Biochemistry, Facutly of Dentistry, 歯学部, 講師 (20112063)
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| Co-Investigator(Kenkyū-buntansha) |
SHIRAI Eiji Department of Biochemistry, Faculty of Dentistry, 歯学部, 助手 (70171030)
SUZUKI Fujio Department of Biochemistry, Faculty of Dentistry, 歯学部, 教授 (40028717)
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| Project Period (FY) |
1986 – 1987
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| Project Status |
Completed (Fiscal Year 1987)
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| Budget Amount *help |
¥6,500,000 (Direct Cost: ¥6,500,000)
Fiscal Year 1987: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1986: ¥5,500,000 (Direct Cost: ¥5,500,000)
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| Keywords | Crtilage-derived Anti-tucmor (CATF) / Cultured Chondrocytes / Established Cell Line / Endothelial Cell / Growth Inhibitor / 血管誘導阻害因子 / 軟骨培養細胞株 |
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| Research Abstract |
1. Purification of cartilage-derived anti-tumor factor (CATF)
CATF was extracted from fetal bovine cartilage with 1 M guanidine hydrochloride and parrially purified by acetone fractionation (45 - 65%), ultrafiltration (Molecular weight; 100-300k) and DEAE-Sepharose CL-6B column chromatography (eluted with 0.3 to 0.35 M NaCl at pH 8.0). The partially purified CATF strongly inhibited the growth of solid sarcoma 180 and B16 melanoma in vivo, the proliferation and DNA synthesis of bovine pulmonary artery endothelial (CPAE) cells in culture and B16 melanoma-induced angiogenesis in chick embryo chorioallantoic membranes (CAM). The specific activity of this partially purified CATF was about 70 times that of crude CATE when determined using its inhibitory action on DNA synthesis in CPAE cells in culture.
2. Production of CATF by rabbit costal chondrocytes in primary culture.
Serum-free medium conditioned by exposure to rabbit costal chondrocytes in primary culture specifically inhibited the proliferation and DBA synthesis of CPAE cells in Culture. The factor in conditioned medium (CM) also inhibited angiohenesis in CAM induced by B16 melanoma and growth of the tumor transplanted onto the CAM. These findings strongly suggest that rabbit costal chondrocytes produce CATF.
3. Establishment of cell lines producing CATF.
A cell line showing some phenotypes of growth cartilage, such as responsiveness to parathyroid hormone and high alkaline phosphatase activity was established from secondary cultures of mouse costal growth cartilage. Moreover, a cell line showing typical cartilage phenotypes was established from human chondrosarcoma. CM obtained from these cells inhibited the proliferation and DNA synthesis of CPAE cells in culture. The inhibition was specific for CPAE cells. Therefore, these cell lines are thought to produce CATF.
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