alexa Synergism between hepatocytes and Kupffer cells in the activation of fat storing cells (perisinusoidal lipocytes).
Gastroenterology

Gastroenterology

Journal of Gastrointestinal & Digestive System

Author(s): Gressner AM, Lotfi S, Gressner G, Haltner E, Kropf J

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Abstract This study concerns the cooperation of hepatocytes (PC) and Kupffer cells (KC) in the activation of rat liver fat storing cells (FSC) in culture. Various dilutions of conditioned media collected from early, serum-free cultures of both cell types were added separately and in combination, either simultaneously or sequentially, to early, non-confluent, primary cultures of FSC maintained under serum-reduced (0.5\% fetal calf serum) conditions to determine the effects on proliferation (incorporations of [3H]thymidine and bromodeoxyuridine [BrdUrd], DNA-content, cell number), transformation and morphology (phase contrast microscopy, immunostainings of desmin and smooth muscle-alpha-actin), on the deposition of fibronectin and laminin and on the formation of 35S sulfated medium proteoglycans. Media of both cell types stimulated cell proliferation in a dose-dependent manner but combined PC- and KC-conditioned media was most potent and increased the incorporation of [3H]thymidine to 4-times above control values. The multiplication stimulatory effects visualized by labeling cell nuclei with BrdUrd and the increase of cell number per culture well were additive. The sequential addition of KC-conditioned medium to FSC preexposed to PC-conditioned medium increased the multiplication of FSC further and in an additive manner. The mitogenic activity of the PC-medium and the enhancing effect of KC-induced FSC proliferation was measured also when PC were damaged by incubation under anoxic conditions during generation of the conditioned medium. This observation indicates the release of the mitogen by membrane damage presumably from a cytoplasmic pool. The PC-medium did not induce either significant morphological changes or transformation of FSC towards myofibroblast-like cells. KC, however, generated transformation of FSC as indicated by more elongated cells with spindle-like cellular extensions and a reduction of retinoid droplets. Both these morphological effects were visible when PC and KC media were added simultaneously. Both media act synergistically on the deposition of fibronectin and laminin in FSC cultures and these components were found to be elevated 2.3 and 2.8-fold, respectively, if the cells were exposed to the combined media. Proteoglycan synthesis was also maximally enhanced if FSC were exposed to PC- and KC-media simultaneously. These findings suggest the involvement of (damaged) hepatocytes in the process of FSC activation. A model of sequential, spatial and time-dependent activation of FSC is suggested where cells in the immediate proximity of hepatocytes are primed to proliferate by a mitogenic signal released by membrane damage presumably from a cytoplasmic pool of injured hepatocytes into the pericellular environment. This non-inflammatory stimulation is followed by secretions of activated Kupffer cells and other inflammatory cell types which further enhance the activation of FSC.(ABSTRACT TRUNCATED AT 400 WORDS)
This article was published in J Hepatol and referenced in Journal of Gastrointestinal & Digestive System

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