[Continuous perfusion of dispersed anterior pituitary cells: a model for LHRH regulation of LH secretion].

ABSTRACT

The anterior pituitary (AP) tissues were removed from adult male Sprague-Dawley rats by decapitation. The dispersed cells with high viability (greater than or equal to 95%) were prepared using trypsin digestion and mechanical dissociation, and then mixed with Bio-Gel P2, packed into the columns, and perfused continuously with M199 medium for more than 24 hours. Different doses of LHRH were administered by 6-min pulses at one hour intervals. A steady and detectable basal LH secretion was present in all columns during the experiment. LHRH stimulation pulse could induce LH secretion rapidly, and repeated LHRH pulses of same dose produced statistically equal LH pulses. The dose-response curve of LH secretion was linear within the range of 1 X 10(-10) to 1 X 10(-7) mol/L LHRH. These results indicate that continuous perfusion system of dispersed AP cells which offers significant advantages over other methodologies provides a very useful in vitro model for studying the mechanisms on LHRH regulation of LH secretion.