Modification of cell perifusion for extended study of hormone release in the rat pituitary.
We use a flow-through cell perifusion system allowing continuous sampling of Luteinizing Hormone (LH) release from excised pituitary tissue. With this system, we are able to sample from viable tissue in primary culture for three days as opposed to a more typical of primary culture of 6-12 hours. Rat pituitary tissue was perifused and challenged with a physiological dose of LHRH. Samples were collected for 3.5 hours on each of 3 consecutive days in a refrigerated fraction collector and assayed for LH concentration by radioimmunoassay. In these experiments we tested two types of media, each with or without fetal calf serum supplementation, for their ability to support pituitary tissue for extended lengths of perifusion. In addition, we broadened the use of aseptic technique, sterile media and equipment to limit microbial contamination. Our results indicate that perifusion of pituitary tissue can be done successfully for 3 or more days at a modest cost, greatly increasing the amount of information that can be collected from each piece of tissue harvested.
Vella
S
S
University of Wisconsin-Whitewater, Department of Biological Sciences, Whitewater, WI, USA.
Gussick
J
J
Woller
M
M
Waechter-Brulla
D
D
D000818:Animals; D001798:Blood Proteins / Q000494:pharmacology; D002470:Cell Survival / Q000187:drug effects / Q000502:physiology; D002478:Cells, Cultured; D003470:Culture Media / Q000737:chemistry / Q000494:pharmacology; D015194:Diffusion Chambers, Culture / Q000295:instrumentation / Q000379:methods*; D007987:Gonadotropin-Releasing Hormone / Q000494:pharmacology; D007260:Infusion Pumps; D007986:Luteinizing Hormone / Q000378:metabolism*; D008297:Male; D010477:Perfusion / Q000295:instrumentation / Q000379:methods*; D010903:Pituitary Gland, Anterior / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D015203:Reproducibility of Results; D013242:Sterilization / Q000295:instrumentation / Q000379:methods
D001798:Blood Proteins; D003470:Culture Media; D007987:Gonadotropin-Releasing Hormone; D007986:Luteinizing Hormone