title: Comparative effects of UW and SLS solutions on concentrative proline uptake in cold preserved rat hepatocytes.
abstract: In previous studies, we have shown that the rate of cell swelling induced by concentrative proline uptake in isolated rat hepatocytes decreased by 50 per cent after only 24 h of cold storage in University of Wisconsin (UW) solution, thereby representing a sensitive marker of alterations in hepatocyte functions after cold preservation and rewarming. We have thus used concentrative proline uptake to compare the capacity of UW and sodium-lactobionate-sucrose (SLS) solutions to maintain such differentiated hepatocyte functions. Isolated rat hepatocytes were kept at 4 degrees C for 4, 10, 24 and 48 h in UW or SLS solutions, and subsequently cultured at 37 degrees C for 1-2 h. Viability was measured by Trypan blue exclusion. After rewarming, cells were subjected to a 10 min administration of 10 mM proline and accumulation of the amino acid was assessed by changes in cell volume as measured by digital analysis of single-cell images obtained under bright-field illumination. Cell viability was reduced gradually and significantly after 0 to 48 h of preservation, and rewarming amplified this effect. However, loss of viability was similar in UW- and SLS-stored cells, as were initial steady-state cell volumes. Proline-induced swelling rate was reduced significantly by 13, 46 and by 57 per cent after 10, 24 and 48 h of preservation in UW solution, respectively. There is no significant difference between SLS- and UW-preserved hepatocyte swelling rates after 10 h and 48 h of cold preservation. However, the decline in the swelling rate of SLS-preserved hepatocytes incubated for 24 h is significantly lower than that of their UW-preserved counterparts. These results show that the SLS solution can preserve differentiated hepatic functions as well as the UW solution does.
authors.affiliation: Département de pharmacologie, Université de Montréal, Québec, Canada.
authors.forename: H
authors.initials: H
authors.lastname: Serrar
authors.forename: L
authors.initials: L
authors.lastname: Musallam
authors.forename: P
authors.initials: P
authors.lastname: Haddad
mesh_terms: D000241:Adenosine / Q000494:pharmacology; D000493:Allopurinol / Q000494:pharmacology; D000818:Animals; D002454:Cell Differentiation; D048429:Cell Size / Q000187:drug effects; D002470:Cell Survival; D002746:Chlorpromazine / Q000494:pharmacology; D015925:Cryopreservation / Q000379:methods*; D004187:Disaccharides / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D005978:Glutathione / Q000494:pharmacology; D007328:Insulin / Q000494:pharmacology; D008099:Liver / Q000166:cytology*; D016031:Liver Transplantation; D008297:Male; D019188:Organ Preservation Solutions; D011392:Proline / Q000378:metabolism*; D011887:Raffinose / Q000494:pharmacology; D051381:Rats; D017207:Rats, Sprague-Dawley; D013395:Sucrose / Q000494:pharmacology / Q000031:analogs & derivatives; D014021:Tissue Preservation / Q000379:methods*
chemical_list: D004187:Disaccharides; D007328:Insulin; D019188:Organ Preservation Solutions; C055206:University of Wisconsin-lactobionate solution; C067074:sodium lactobionate sucrose solution; D013395:Sucrose; D000493:Allopurinol; D011392:Proline; D005978:Glutathione; D000241:Adenosine; D011887:Raffinose; D002746:Chlorpromazine