engleski

Development and characterisation of rat anterior pituitary primary cell culture - biochemical approach

We describe an isolation method that enables us to study the regulation of pituitary hormones secretion within anterior pituitary cell cultures. Tissues in vitro systems have many limitations, such as: the requirement of many donor animals for relatively few experiments, poor reproducibility and short culture life. New model consisting of dispersed pituitary cells provides multiple treatment capability, and adaptability. We used mechanical disruption method and short-term enzymatic dispersion (collagenase). Pituitary cel suspensions attached quickly (within 24 hours) and quantitatively (85%) to plastic surfaces in medium M-199 (pH 7,6) containing 10% foetal calf serum. Such attached anterior pituitary cells discovered the subpopulation of different cell types that were detected by electron microscopy of the culture. After prolongrd cultivation the cells have established colonies and monolayer structure. The optimal density of the cells was between 1-5x10^5 cells/mL. Such culture of pituitary cells 24 hours after addition of 4-^14C-testosterone converted it into 5alpha-dihydrotestosterone. Pituitary cells after isolation from tissue and culturing showed comparable activity with tissue culture experiments made previously. The high density plating procedure described here provides greater opportunity for cell-to-cell interaction and it can be useful model for evaluating the role of intercellular communication within the tissue.

anterior pituitary; cell culture; prolonged cell cultivation

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