Background. University of Wisconsin solution (UWS) has been successfully used for liver transplantation and may be beneficial for hypothermic cardiac storage. The addition of glucose may enhance myocardial preservation. Methods and Results. Cultured human ventricular myocytes (eight dishes per group) were stored at 0-degrees-C for 12 hours in either unmodified UWS or UWS with glucose (1, 3, 10, 30, or 100 mmol/l). Cells were assayed for protein by spectrofluorometry and adenine nucleotides by high performance liquid chromatography after storage. Protein recovery, adenosine triphosphate (ATP), adenosine diphosphate (ADP), and total adenine nucleotides (ATP+ADP+AMP) were all depleted after storage (p<0.0001 by ANOVA). Protein recovery (p<0.005), ATP (p<0.05), and ADP (p<0.05) were increased with glucose administration compared with unmodified UWS. Improvement was maximal using 30 mmol/l (protein, 0 mmol/l =0.48+/-0.14 and 30 mmol/l=0.65+/-0.11 mg per dish; ATP, 0 mmol/l=3.08+/-0.63 and 30 mmol/l =4.32+/-0.90 nmol/mg protein; ADP, 0 mmol/l=3.76+/-0.80 and 30 mmol/l=4.63+/-0.38 nmol/mg protein, mean+/-SD). Total adenine nucleotides tended to increase at any glucose concentration (p=0.07 by ANOVA) and were significantly better with 30 mmol/l glucose (0 mmol/l=7.61+/-1.58 and 30 mmol/l =9.62+/-1.08 nmol/mg protein). Conclusions. Increasing the glucose concentration from 0 to 30 mmol/l improved adenine nucleotide and cellular protein preservation in this in vitro assessment.