Preservation studies using acinar cell cultures of the pancreas: stimulation of amylase/lipase release before and after hypoxic stress.
In clinical pancreas transplantation, postischemic (i. e. postpreservation) transplant pancreatitis is a major problem in some cases but also an interesting model of pancreatitis. In this study, the effect of simulated organ preservation of isolated acinar cells was evaluated as regards enzyme release (basic and cerulein-stimulated), using common preservation solutions as the incubation media. Primary pancreas acinar cell cultures were isolated from the pancreas of male Wistar rats using the modified method of Amsterdam and Jamieson. After resting the cells in culture flasks for 1 week, monolayer cultures were obtained. The basic enzyme release of amylase and lipase was measured as well as the effect of stimulation with cerulein (10(-8) M) and the effect of replacing the medium (without changing the consistence or temperature of the medium). In a second step, the cultures were incubated under conditions of cold hypoxia for 6 h (4 degrees C, P O2 < 0.1 mm Hg) using Krebs-Henseleit solution (KH), Euro-Collins solution (EC), HTK solution of Bretschneider (HTK) or University of Wisconsin solution (UW) as the incubation solution. After 6 h, the media were changed to warm normoxic KH, and a second stimulation test with cerulein was performed. The native microstructure of the cultures was observed as well. Enzyme release was elevated by a factor of 5 by stimulating the acinar cells with cerulein as well as by changing the medium in the experiments prior to the hypoxic incubation. After hypoxic incubation and change to KH, the morphology of the cultures was excellent, while the basic enzyme release was on a very low level, no matter which preservation solution was used during cold hypoxia. Stimulation with cerulein caused only minimal elevation of enzyme release during an observation period of 60 min. These observations show that cold storage in preservation solution provides maintenance of the cell morphology and sufficient down-regulation of enzyme release of pancreatic acinar cells. Thus, acinar cells alone do not seem to be the pacemaker of pancreatitis after organ preservation. The presented experimental model will be the subject of extended evaluation in the future.
Department of General Surgery, University of Tübingen, Germany.
R
R
Viebahn
B
B
Metzdorf
I
I
Besenthal
D000681:Amylases / Q000378:metabolism*; D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods; D015687:Cell Hypoxia / Q000502:physiology*; D002478:Cells, Cultured; D007700:Kinetics; D008049:Lipase / Q000378:metabolism*; D008297:Male; D019188:Organ Preservation Solutions; D010179:Pancreas / Q000166:cytology* / Q000201:enzymology*; D051381:Rats; D017208:Rats, Wistar; D013997:Time Factors; D014021:Tissue Preservation / Q000379:methods*
D019188:Organ Preservation Solutions; D008049:Lipase; D000681:Amylases