Cardiac myocyte functional and biochemical changes after hypothermic preservation in vitro. Protective effects of storage solutions.
In this study, we evaluated cardiac myocyte viability and function under hypothermic conditions with four types of storage solutions, saline solution, Euro-Collins solution, University of Wisconsin solution, and MCDB 107 medium. Cardiac myocytes were isolated from neonatal rat ventricles by collagenase dispersion and cultured for 4 days with MCDB 107 medium. A total of 12.5 x 10(5) myocytes per culture dish was used and the myocytes were incubated at 4 degrees C for 6, 12, 18, and 24 hours in the various storage solutions. After each incubation time, creatine kinase and lactate dehydrogenase were measured in the storage solutions. The myocytes were then incubated for 24 hours at 37 degrees C to evaluate the recovery of the myocyte beating rate. In the MCDB 107 group (n = 7), the recovery ratio of myocyte beating rate was complete by 12 hours, then decreased to 44.8% of control (beating rate before hypothermic incubation) at 24 hours. The saline, Euro-Collins, and University of Wisconsin groups (n = 7 each) had significantly lower recovery ratios than the MCDB 107 group (at 12 hours: 61.0%, 32.2%, and 48.9%; at 18 hours: 0.0%, 5.5%, and 15.1% of control, respectively). Release of creatine kinase and lactate dehydrogenase in the MCDB 107 group gradually increased and at 24 hours was 143.2 mIU/flask and 486.2 mIU/flask, respectively. However, the saline and University of Wisconsin groups had significantly increased creatine kinase and lactate dehydrogenase values at 24 hours (creatine kinase: 334.6 and 319.6 mIU/flask; lactate dehydrogenase: 821.6 and 654.4 mIU/flask, respectively). The Euro-Collins group showed the greatest increase in both markers (creatine kinase: 1587.5, lactate dehydrogenase: 2106.9 mIU/flask). In summary, saline and University of Wisconsin solutions showed a beneficial effect on recovery of myocyte viability at 12 hours compared with Euro-Collins solution, however MCDB 107 medium had the best overall protective effect on cultured myocytes. Accordingly, alternate hypothermic storage solutions, such as cell-culture medium, may have protective characteristics that are suitable for cardiac preservation.
Orita
H
H
Second Department of Surgery, Yamagata University School of Medicine, Japan.
Fukasawa
M
M
Hirooka
S
S
Uchino
H
H
Fukui
K
K
Kohi
M
M
Washio
M
M
D000241:Adenosine; D000493:Allopurinol; D000596:Amino Acids; D000818:Animals; D000831:Animals, Newborn; D002314:Cardioplegic Solutions; D002470:Cell Survival; D002478:Cells, Cultured; D003080:Cold Temperature; D003402:Creatine Kinase / Q000378:metabolism; D005947:Glucose; D005978:Glutathione; D006982:Hypertonic Solutions; D007328:Insulin; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008297:Male; D009200:Myocardial Contraction; D009206:Myocardium / Q000166:cytology / Q000378:metabolism*; D019188:Organ Preservation Solutions; D011887:Raffinose; D051381:Rats; D017208:Rats, Wistar; D012965:Sodium Chloride; D014021:Tissue Preservation; D014168:Transferrin; D014815:Vitamins
D000596:Amino Acids; D002314:Cardioplegic Solutions; C037096:Euro-Collins' solution; D006982:Hypertonic Solutions; D007328:Insulin; C085252:MCDB 107; D019188:Organ Preservation Solutions; D014168:Transferrin; C055206:University of Wisconsin-lactobionate solution; D014815:Vitamins; D012965:Sodium Chloride; D000493:Allopurinol; D007770:L-Lactate Dehydrogenase; D003402:Creatine Kinase; D005978:Glutathione; D005947:Glucose; D000241:Adenosine; D011887:Raffinose