Effect of aging on GHRF-induced growth hormone release from anterior pituitary cells in primary culture.
Five criteria were developed to validate the primary cell culture model for comparison of GRF-induced release of growth hormone in pituitary tissue from aging animals. Pituitaries from young (5-mo), middle-aged (14-mo), and old (24-mo) male Fischer 344 rats were dispersed using either trypsin/trypsin inhibitor or dispase and compared with respect to the number of pituitary cells recovered, cell viability, 3H-leucine incorporation into total protein, time course for recovery of optimal response to GRF, and the dose-relationship for GRF-induced release of growth hormone 2, 4, and 6 days after dispersal. Results indicated that direct comparison of cellular responses between tissues from young, middle-aged, and old rats in primary cell culture is confounded by variations in time for recovery of optimal responses, the effects of the enzymes used for dispersal, and the methods used to express the data.
Spik
K W
KW
Bowman Gray School of Medicine, Wake Forest University.
Boyd
R L
RL
Sonntag
W E
WE
D000375:Aging / Q000378:metabolism*; D000818:Animals; D002452:Cell Count; D002478:Cells, Cultured; D010450:Endopeptidases; D013006:Growth Hormone / Q000493:pharmacokinetics*; D013007:Growth Hormone-Releasing Hormone / Q000494:pharmacology*; D007930:Leucine / Q000378:metabolism; D008297:Male; D010903:Pituitary Gland, Anterior / Q000378:metabolism*; D011506:Proteins / Q000378:metabolism; D051381:Rats; D011916:Rats, Inbred F344; D014316:Tritium; D014357:Trypsin
D011506:Proteins; D014316:Tritium; D013006:Growth Hormone; D013007:Growth Hormone-Releasing Hormone; D010450:Endopeptidases; D014357:Trypsin; C023139:dispase; D007930:Leucine