Monoclonal Antibodies against Pig Ovarian Follicular Granulosa Cells Induce Apoptotic Cell Death in Cultured Granulosa Cells

Journal of Veterinary Medical Science

Akira MYOUMOTO²  Yoshimi NIWANO¹  Kazuhiro SAKAMAKI⁵  Noboru MANABE⁴  Yuzuru IMAI²  Miki SUGIMOTO²  Yoshihiro KIMURA²  Hajime MIYAMOTO²  Manabu FUKUMOTO³  Yoshinori OKAMURA³ 

[1] Institute for Life Science Research, Nihon Nohyaku Co., Ltd.;Department of Animal Sciences, Kyoto University;Department of Pathology I, School of Medicine, Kyoto University;Unit of Anatomy and Cell Biology, Department of Animal Sciences, Kyoto University;Institute for Virus Research, Kyoto University

关键词: apoptosis;    follicular granulosa cell;    monoclonal antibody;    swine ovary;   

DOI  :  10.1292/jvms.59.641

学科分类：兽医学

来源: Japanese Society of Veterinary Science

【 摘 要 】

References(44)Cited-By(11)Two monoclonal antibodies capable of inducing granulosa cell apoptosis were produced against granulosa cells prepared from antral follicles of pig ovaries. The healthy follicles, 4-5 mm in diameter, were dissected from the ovaries of gilts, and then granulosa cells were isolated. BALB/c female mice were immunized with the isolated granulosa cells. Antibodies against the granulosa cells were detected by immunofluorescent staining using frozen ovarian sections. The isolated spleen cells prepared from immunized mice producing antibodies against the granulosa cells were fused with Sp2/O-Ag14 mouse myeloma cells by standard hybridization techniques. Two hybridoma clones, PFG-1 and PFG-2, which produced specific IgM antibodies against granulosa cells were selected. Western blotting analysis revealed that PFG-1 and PFG-2 antibodies specifically recognized cell-membrane proteins with molecular weights of 55 and 70 kD and isoelectric points of 5.9 and 5.4, respectively. The monoclonal antibodies immunohistochemically reacted with granulosa cells of healthy follicles. When the isolated granulosa cells prepared from healthy follicles were cultured in medium containing 0.1 or 10 μg/m l PFG-1 or PFG-2 antibodies, respectively, the cells underwent apoptosis as determined by nuclear morphology, DNA electrophoresis and flow cytometric analysis. In conclusion, these two monoclonal antibodies against granulosa cells have cell-killing activity in cultured granulosa cells.

【 授权许可】

Unknown