Importance of pH in resuspension media on viability of hepatocytes preserved in University of Wisconsin solution.
The effect of different pH of resuspension media on the viability of hepatocytes preserved (for 96 h at 4 degrees C) in University of Wisconsin solution (UW solution) was analyzed. After this cold resuspension media storage, we evaluated the rewarming step (incubation time 120 min at 37 degrees C) using different pH levels (6.80, 7.00, 7.20, and 7.40). Cell viability assessed by trypan blue exclusion (TBE) showed a significant difference (p < 0.05) for cells incubated at pH = 7.20. For instance, TBE expressed as percent of change was 78.1 +/- 1.4 compared with cells tested at other pH (pH = 6.80, TBE = 44.2 +/- 9.5; pH = 7.00, TBE = 66.5 +/- 1.1 and pH = 7.40, TBE = 62.0 +/- 1.4). We also evaluated the capacity of these cells both to maintain potassium content (0.509 +/- 0.230 microEq. K+/10(6) cells) and to synthesize urea (5.36 +/- 1.81 mumol Urea/10(6) cells). These results were compared with those obtained from freshly isolated non preserved hepatocytes (0.518 +/- 0.060 microEq. K+/10(6) cells and 5.91 +/- 0.43 mumol Urea/10(6) cells). The results show that viability is pH dependent and suggest that when resuspension media were used, the viability of hepatocytes was improved after 96 h of cold storage.
Mamprin
M E
ME
Departamento Farmacia, Facultad de Cs, Bioquímicas y Farmacéuticas, Universidad Nacional de Rosario, Argentina.
Rodríguez
J V
JV
Guibert
E E
EE
D000241:Adenosine; D000493:Allopurinol; D000818:Animals; D002452:Cell Count; D002470:Cell Survival; D017690:Cell Transplantation; D002478:Cells, Cultured; D005978:Glutathione; D006863:Hydrogen-Ion Concentration; D007328:Insulin; D008099:Liver / Q000166:cytology*; D008297:Male; D019188:Organ Preservation Solutions; D011887:Raffinose; D051381:Rats; D017208:Rats, Wistar; D014021:Tissue Preservation
D007328:Insulin; D019188:Organ Preservation Solutions; C055206:University of Wisconsin-lactobionate solution; D000493:Allopurinol; D005978:Glutathione; D000241:Adenosine; D011887:Raffinose