Zechner-Krpan V, Kniewald J, Kniewald Z
Faculty of Food Technology and Biotechnology, University of Zagreb, Croatia.
Arh Hig Rada Toksikol. 1998 Sep;49(3):219-29.
High density plating procedure was used to evaluate the effect of atrazine on anterior pituitary cells of rats in monolayer culture. Collagenase-dispersed pituitary cells plated in suspension with medium-199 and 10% foetal calf serum attached quantitatively to plastic surfaces within 24 hours. Electron microscopy showed subpopulations of different cell types. After prolonged cultivation, most cells established small colonies with extensive contacts among them. Cell-to-cell formation of aggregates was significant and the colonies manifested morphological changes. The cells retained their enzymatic activity, converting testosterone into 5 alpha-dihydrotestosterone by enzyme 5 alpha-reductase. Immunohistochemical techniques facilitated differentiation of gonadotrophs producing follicle stimulating hormone (FSH) and luteinising hormone (LH). Atrazine in concentrations of 5 to 50 micrograms/ml of medium was associated with a significant reduction in the number of viable cells within 72 hours. The results suggest that the pituitary cell culture may prove useful in toxicological testing of various toxic compounds and reduce or replace in vivo animal experiments.
采用高密度接种程序评估莠去津对单层培养的大鼠垂体前叶细胞的影响。用胶原酶分散的垂体细胞与199培养基和10%胎牛血清一起悬浮接种,在24小时内定量附着于塑料表面。电子显微镜显示了不同细胞类型的亚群。长时间培养后,大多数细胞形成了小菌落,它们之间有广泛的接触。细胞间聚集的形成很显著,菌落表现出形态变化。这些细胞保留了它们的酶活性,通过5α-还原酶将睾酮转化为5α-双氢睾酮。免疫组织化学技术有助于区分产生促卵泡激素(FSH)和促黄体生成素(LH)的促性腺激素细胞。培养基中浓度为5至50微克/毫升的莠去津与72小时内活细胞数量的显著减少有关。结果表明,垂体细胞培养可能在各种有毒化合物的毒理学测试中证明是有用的,并减少或取代体内动物实验。