engleski

Application of immunohistochemistry for the study of pituitary gonadotropins

The aim of this study was to investigate the application of male rat pituitary primary cell culture in toxicological studies. The plating procedure for pituitary cells was successfully applied to perform the atrazine influence on pituitary cells in culture. Collagenase-dispersed pituitary cell suspensions attached within 24 hrs on plastic surfaces when plated in medium-199 (M-199) containing 10% fetal calf serum (FCS). Anterior pituitary cells attached in this way developed the subpopulations of different cell types, which were detected by electron microscopy of the culture. After prolonged cultivation most of the cells established small colonies and contacts among cells appeared abundant. The significant cell-to-cell formation of aggregates was evident, as well as the morphological changes within the colony. The pituitary cells in culture retained their enzymatic activity of converting testosterone into 5a-dihydrotestosterone (5a-DHT) by enzyme 5a-reductase. By introducing the immunohistochemical evaluation into the cell culture of pituitary gland, it was possible to differentiate on the monolayer of pituitary primary culture the cells responsible for follicle stimulating hormone (FSH) or luteinizing hormone (LH) secreting. Herbicide atrazine, was present in the culture of pituitary cells in concentration from 5 to 50 mg per ml of medium, and after 24 or 48 hrs significant reduction of the number of viable cells were detected. Our results showed that the pituitary cells in culture could be also successfully used in the testing of toxic compounds, in order to reduce or replace the experiments with the animals.

pituitary cells; primary cell culture; gonadotropic hormones; immunohistochemistry; toxicology

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