Mild hypothermic preservation for transport purposes of the AMC bioartificial liver charged with porcine hepatocytes.
BACKGROUND
Preservation conditions play a crucial role during transport of a bioartificial liver (BAL) from the laboratory to the hospital. We assessed the possibility to preserve the AMC-BAL loaded with freshly isolated porcine hepatocytes at mild hypothermic temperatures.


METHODS
Two laboratory-scale AMC-bioreactors were loaded with 1 billion freshly isolated porcine hepatocytes per experiment (n=6). Bioreactors in the control group were kept for three days at 37 degrees C. Bioreactors in the transport group were kept at 37 degrees C during day 1, at 15 degrees C during day 2, and again at 37 degrees C during day 3. In addition, long-term mild hypothermic preservation periods of 45 and 110 hr at 15 degrees C and 26 degrees C, respectively, were assessed. The effect of mild hypothermic preservation on hepatocytes inside the bioreactors was tested by determination of cell damage parameters, as well as metabolic and hepatocyte-specific functions.


RESULTS
A 24-hour period of mild hypothermic preservation did not reduce any hepatocyte-specific function. LDH release was significantly higher only at day 2. Albumin production at day 2 and lidocaine elimination at day 3 were significantly higher with glucose consumption and lactate production being significantly lower at both test days. Long-term mild hypothermic preservation had a drastic negative effect on cellular viability and hepatocyte-specific function.


CONCLUSIONS
Mild hypothermic preservation at temperatures as low as 15 degrees C and for a duration of 24 hr is a feasible method to preserve BAL systems loaded with freshly isolated porcine liver cells and will simplify the logistics of BAL transport from the laboratory to the hospital.
Poyck
Paul P C
PP
Department of Surgery, Academic Medical Center, University of Amsterdam, Amsterdam, the Netherlands.
Hoekstra
Ruurdtje
R
van Wijk
Albert C W A
AC
ten Bloemendaal
Lysbeth
L
Chamuleau
Robert A F M
RA
van Gulik
Thomas M
TM
D000418:Albumins / Q000096:biosynthesis; D000818:Animals; D002470:Cell Survival; D003080:Cold Temperature; D005260:Female; D005947:Glucose / Q000378:metabolism; D022781:Hepatocytes / Q000378:metabolism / Q000502:physiology; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D019344:Lactic Acid / Q000096:biosynthesis; D008012:Lidocaine / Q000493:pharmacokinetics; D019164:Liver, Artificial; D013552:Swine; D013997:Time Factors; D014021:Tissue Preservation; D014186:Transportation
D000418:Albumins; D019344:Lactic Acid; D008012:Lidocaine; D007770:L-Lactate Dehydrogenase; D005947:Glucose