P. Olinga et al., INFLUENCE OF 48 HOURS OF COLD-STORAGE IN UNIVERSITY-OF-WISCONSIN ORGAN PRESERVATION SOLUTION ON METABOLIC CAPACITY OF RAT HEPATOCYTES, Journal of hepatology, 27(4), 1997, pp. 738-743
Background/Aims: Suspensions of isolated hepatocytes are a valuable to
ol to study liver functions, For optimal use of the isolated hepatocyt
es, methods are needed to preserve the hepatocytes while maintaining t
heir viability, metabolic and transport functions, Until now little ha
s been known about the maintenance of the drug metabolism capacity and
energy state, measured by the so-called energy charge (ATP+1/2ADP)/(A
TP+ADP+AMP), in hepatocytes after storage in University of Wisconsin c
old storage solution (UW), Consequently, we investigated whether UW, o
riginally designed to preserve organs for transplantation, was suitabl
e for preservation of isolated rat hepatocytes with respect to the mai
ntenance of drug metabolism and levels of energy-rich substrates. Meth
ods: Viability of the isolated rat hepatocytes was determined by trypa
n blue exclusion, ATP content and energy charge after 24 and 48 h of s
torage in UW at 0 degrees C, Phase I and II metabolic functions of the
cells were studied by measuring the cytochrome P450 content and the m
etabolic rate of lidocaine and 7-ethoxycoumarin. Results: During 48 h
of storage of hepatocytes in UW both phase I and phase II metabolism a
re preserved at control levels, After storage, the viability of the he
patocytes was not changed significantly, and the cells maintained prop
er cellular ATP content and overall energy charge. Conclusions: These
results imply that hepatocytes from a single isolation can be stored i
n UW solution and used for metabolism experiments for 3 consecutive da
ys, allowing a reduction in the use of experimental animals.