Addition of L-carnitine to additive solution-suspended red cells stored at 4 degrees C reduces in vitro hemolysis and improves in vivo viability.
BACKGROUND
The role of L-carnitine (LC) as the requisite carrier of long-chain fatty acids into mitochondria is well established. Human red cells (RBCs), which lack mitochondria, possess a substantial amount of LC and its esters. In addition, carnitine palmitoyl transferase, an enzyme that catalyzes the reversible transfer of the acyl moiety from acyl-coenzyme A to LC is found in RBCs. It has recently been shown that LC and carnitine palmitoyl transferase play a major role in modulating the pathway for the turnover of membrane phospholipid fatty acids in intact human RBCs, and that LC improved the membrane stability of RBCs subjected to high shear stress. RBC membrane lesions occur during storage at 4 degrees C; this study investigated whether the addition of LC (5 mM) to a standard RBC preservative solution (AS-3) affected cellular integrity with 42 days' storage.


METHODS
A paired (n = 10) crossover design was used for RBCs stored in AS-3 with and without LC. Both in vitro RBC properties reflective of metabolic and membrane integrity and in vivo measures of cell viability (24-hour percentage of recovery and circulating lifespan) were measured at the end of the storage. In addition, the turnover of membrane phospholipid and long-chain acylcarnitine fatty acids and the carnitine content of control and LC-stored RBCs were measured.


RESULTS
It was shown that LC was irreversibly taken up by RBCs during storage, with a fourfold increase at 42 days. Furthermore, as found by the use of radiolabeled palmitate, the stored RBCs were capable of generating long-chain acylcarnitine. The uptake of LC during storage was associated with less hemolysis and higher RBC ATP levels and by a significantly greater in vivo viability for LC-stored RBCs than for control-stored RBCs: a mean 24-hour percentage of recovery of 83.9 +/- 5.0 vs. 80.1 +/- 6.0 percent and a mean lifespan of 96 +/- 11 vs. 86 +/- 14 days, respectively (p < 0.05).


CONCLUSIONS
A beneficial effect of the addition of LC to RBCs stored at 4 degrees C was evident. This effect may be related to both biophysical and metabolic actions on the cell membrane.
Arduini
A
A
Department of Sciences, University G. D'Annunzio, Pescara, Italy.
Holme
S
S
Sweeney
J D
JD
Dottori
S
S
Sciarroni
A F
AF
Calvani
M
M
D000225:Adenine / Q000494:pharmacology; D001793:Blood Preservation / Q000379:methods*; D002331:Carnitine / Q000494:pharmacology*; D002951:Citrates / Q000494:pharmacology; D015925:Cryopreservation; D004905:Erythrocyte Aging / Q000187:drug effects*; D004912:Erythrocytes; D005947:Glucose / Q000494:pharmacology; D006461:Hemolysis / Q000187:drug effects*; D006801:Humans; D010710:Phosphates / Q000494:pharmacology; D012965:Sodium Chloride / Q000494:pharmacology
C058647:AS-3 preservative solution; D002951:Citrates; D010710:Phosphates; D012965:Sodium Chloride; D005947:Glucose; D000225:Adenine; D002331:Carnitine