Abstract
Abstract
KupVer cells (KC) are the resident macrophages of the liver and represent about 80% of the total Wxed macrophage population. They
are involved in disease states such as endotoxin shock, alcoholic liver diseases and other toxic-induced liver injury. They release physiologically
active substances such as eicosanoids and inXammatory cytokines (IL-1, IL-6, TNF�), and produce free radical ...
Abstract
Abstract
KupVer cells (KC) are the resident macrophages of the liver and represent about 80% of the total Wxed macrophage population. They
are involved in disease states such as endotoxin shock, alcoholic liver diseases and other toxic-induced liver injury. They release physiologically
active substances such as eicosanoids and inXammatory cytokines (IL-1, IL-6, TNF�), and produce free radical species. Thus,
KC are attractive targets for anti-inXammatory therapies and potential candidates responsible for diVerences in inXammation in liver disease
seen between diVerent individuals. However, to perform parallel in vitro experiments with KC from diVerent donors a suitable
method for conservation of KC would be necessary. Therefore, the present study evaluated, whether rat and human KC can be frozen,
stored and recovered without losing their functional integrity. Rat and human KC were isolated and either cultured under standard conditions
(fresh KC) or cryopreserved in special freezing medium (cryopreserved KC). At least 24 h later, cryopreserved KC were thawed,
brought into suspension and seeded in the same density as fresh cells for subsequent experiments. Viability of cultured KC was analyzed
by trypan blue exclusion. LPS (or PBS as control) stimulation was performed at diVerent time points and cytokine release was analyzed
with IL-6 and TNF� ELISAs, respectively. Phagocytic capacity was investigated by using a speciWc phagocytosis assay and FACS analysis.
The recovery rate after thawing was around 57% for rat and around 65% for human cryopreserved KC. The results indicate, that KC
can successfully be cryopreserved with an adequate recovery rate of viable cells. The properties of fresh and frozen KC can also be compared
after thawing. Freshly isolated and cryopreserved cultured KC showed near-normal morphology and did not diVer in the cultivation
proWles over a period of 72 h. One to three days after seeding, frozen rat or human KC also retained inducible functions such as the
production of TNF� or IL-6 after LPS challenge. Finally, regardless if they were cryopreserved or not, no diVerences in the phagocytic
activities of the cells were obtained. Taken together, it is concluded that cryopreservation of KC does not change the physiological characteristics
of the cells in vitro. Therefore, the method used here for cryopreservation of especially human KC allows the accumulation of
KC from several donors for parallel in vitro experiments.
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