BIOLOGY OF THE MALE REPRODUCTIVE TRACT
7:30 AM-10:00 AM
(573) ESTABLISHMENT AND CHARACTERIZATION OF CELL LINES CLONED FROM SEMINAL VESICLES OF p53-DEFICIENT MICE.
Kanazawa, Takuya1, Aizawa, Shinichi2, Tomooka, Yasuhiro3, 1 Div of Applied Life Sciences, Ami, Ibaraki, Japan2 Inst of Embryology and Genetics, Kumamoto, Japan3 Dept of Science and Technology, Noda, Japan
ABSTRACT- Seminal vesicles are one of the accessory glands of the male reproductive tract, and secrete components of seminal fluid, which play an important role in maturation and motility of the spermatozoa. Development and function of seminal vesicles are controlled by gonadotropins and sex steroids. However insights into molecular mechanisms of these are limited, due to lack of suitable cell culture systems. Recently, deficiency of p53 has been reported to result in enhanced proliferative potential of cells in culture, which should help in establishing cell lines from seminal vesicles. We have therefore attempted to establish cell lines from seminal vesicles of p53-deficient mice, to provide a model system for studying molecular mechanisms involved in growth and function of the glandular epithelial cells. Cells were dissociated from seminal vesicles of p53-knockout mice, and cultured in monolayer with fetal bovine serum (FBS)-supplemented medium. After cloning, six cell lines were established. These cell lines, designated as p53dmSV-1, -2, -3, -4, -5 and -6, were all positive for keratin and negative for vimentin, which is characteristic of epithelial cells. Growth profiles in monolayer culture with 10% FBS-supplemented medium indicated that population doubling times of these cells varied from 13 h to 60 h. These cell lines differ in morphology and responsiveness to growth factors (insulin, EGF, FGF-2) both in monolayer culture and in collagen gel matrix culture with 3% FBS in medium. SV-4 cells, in particular, responded to dihydroxytestosterone with growth promotion in collagen gel matrix culture and formation of domes, semi-spherical multicellular structures, in monolayer culture. These results indicate that 1) p53-deficient mice serve as good sources for establishing cell lines, 2) these clonal cell lines would be useful for studying growth control of seminal vesicle epithelial cells by sex steroids and local growth factors. (Grant-in-Aid for Scientific Research (C) to T. K. and Grant-in-Aid for Scientific Research to T. Y., both from The Ministry of Education, Culture, Sports, Science and Technology of Japan)
KEY WORDS: seminal vesicles, p53-deficient mouse, clonal cell, dihydroxytestosterone